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    Structured Review

    atcc lmh cells
    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. <t>LMH</t> <t>cells</t> transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Lmh Cells, supplied by atcc, used in various techniques. Bioz Stars score: 95/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    2) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    3) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    4) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    5) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    6) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    7) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    8) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    9) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    10) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    11) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    12) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    13) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    14) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    15) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    16) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    17) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    18) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    19) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    20) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    21) Product Images from "Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes"

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2022.1003262

    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
    Figure Legend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Techniques Used: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.
    Figure Legend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Techniques Used: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.
    Figure Legend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Techniques Used: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.
    Figure Legend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Techniques Used: Neutralization, Western Blot, Activity Assay, Infection

    22) Product Images from "Elagolix is porphyrogenic and may induce porphyric attacks in patients with the acute hepatic porphyrias"

    Article Title: Elagolix is porphyrogenic and may induce porphyric attacks in patients with the acute hepatic porphyrias

    Journal: Molecular Genetics and Metabolism Reports

    doi: 10.1016/j.ymgmr.2022.100915

    Porphyrogenicity and cytotoxicity of elagolix in LMH cells. (a) The fluorescence readings produced by 0.314 mM acetaminophen and 0.314 mM AIA, the negative and positive controls, respectively, clearly indicate porphyrogenicity and non-porphyrogenicity, respectively. Higher concentrations of ALA produced readings greater than the readings produced by AIA while all concentrations of acetaminophen produced readings below the readings produced by AIA. Elagolix produced readings similar to the readings by AIA. DFO is an iron chelator that prevents the conversion of the fluorescent intermediate protoporphyrin to heme, increasing the amount of fluorescent protoporphyrin and mimicking the effects of the porphyrias. (b) The drug screening assay was completed in parallel for each compound to assess for cytotoxicity via the ATPLite cytotoxicity assay. CC 50 values were calculated using the software, GraphPad Prism 8. Data are presented as mean values ± SEM of 3 independent replicates. All results are representative of three independent experiments. A two-sided Student's t- test was used to assess the statistical significance between the two ratios.
    Figure Legend Snippet: Porphyrogenicity and cytotoxicity of elagolix in LMH cells. (a) The fluorescence readings produced by 0.314 mM acetaminophen and 0.314 mM AIA, the negative and positive controls, respectively, clearly indicate porphyrogenicity and non-porphyrogenicity, respectively. Higher concentrations of ALA produced readings greater than the readings produced by AIA while all concentrations of acetaminophen produced readings below the readings produced by AIA. Elagolix produced readings similar to the readings by AIA. DFO is an iron chelator that prevents the conversion of the fluorescent intermediate protoporphyrin to heme, increasing the amount of fluorescent protoporphyrin and mimicking the effects of the porphyrias. (b) The drug screening assay was completed in parallel for each compound to assess for cytotoxicity via the ATPLite cytotoxicity assay. CC 50 values were calculated using the software, GraphPad Prism 8. Data are presented as mean values ± SEM of 3 independent replicates. All results are representative of three independent experiments. A two-sided Student's t- test was used to assess the statistical significance between the two ratios.

    Techniques Used: Fluorescence, Produced, Screening Assay, Cytotoxicity Assay, Software

    23) Product Images from "Eslicarbazepine acetate is porphyrogenic and should be used with caution in patients with the acute hepatic porphyrias"

    Article Title: Eslicarbazepine acetate is porphyrogenic and should be used with caution in patients with the acute hepatic porphyrias

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2022.953961

    Cytotoxicity of eslicarbazepine acetate and other compounds in LMH cells. The ATPLite cytotoxicity assay was completed in parallel with the drug screening assays. CC 50 values were calculated using the software, GraphPad Prism 8. All data are presented as mean values ± SEM of three independent replicates. All results are representative of three independent experiments.
    Figure Legend Snippet: Cytotoxicity of eslicarbazepine acetate and other compounds in LMH cells. The ATPLite cytotoxicity assay was completed in parallel with the drug screening assays. CC 50 values were calculated using the software, GraphPad Prism 8. All data are presented as mean values ± SEM of three independent replicates. All results are representative of three independent experiments.

    Techniques Used: Cytotoxicity Assay, Software

    Porphyrogenicity of eslicarbazepine acetate and other compounds in LMH cells. Deferoxamine (DFO) is an iron chelator that prevents the conversion of the fluorescent intermediate, protoporphyrin, into heme. DFO is added in all experimental groups to mimic the conditions of the AHPs and to increase fluorescence measurements read by the fluorospectrometer. 0.314 mM aspirin and allyl isopropyl acetamide were negative and positive controls, respectively, and were added in all experiments to distinguish porphyrogenicity from non-porphyrogenicity in selected compounds. Higher concentrations of carbamazepine, eslicarbazepine acetate, and phenobarbital produced fluorescence readings greater than the readings produced by 0.314 mM AIA while all concentrations of aspirin produced readings below the readings produced by AIA. All data are presented as mean values ±SEM of three independent replicates. All results are representative of three independent experiments. A two-sided Student’s t -test was used to assess statistical significance.
    Figure Legend Snippet: Porphyrogenicity of eslicarbazepine acetate and other compounds in LMH cells. Deferoxamine (DFO) is an iron chelator that prevents the conversion of the fluorescent intermediate, protoporphyrin, into heme. DFO is added in all experimental groups to mimic the conditions of the AHPs and to increase fluorescence measurements read by the fluorospectrometer. 0.314 mM aspirin and allyl isopropyl acetamide were negative and positive controls, respectively, and were added in all experiments to distinguish porphyrogenicity from non-porphyrogenicity in selected compounds. Higher concentrations of carbamazepine, eslicarbazepine acetate, and phenobarbital produced fluorescence readings greater than the readings produced by 0.314 mM AIA while all concentrations of aspirin produced readings below the readings produced by AIA. All data are presented as mean values ±SEM of three independent replicates. All results are representative of three independent experiments. A two-sided Student’s t -test was used to assess statistical significance.

    Techniques Used: Fluorescence, Produced

    24) Product Images from "Assessment of porphyrogenicity of drugs and chemicals in selected hepatic cell culture models through a fluorescence‐based screening assay. Assessment of porphyrogenicity of drugs and chemicals in selected hepatic cell culture models through a fluorescence‐based screening assay"

    Article Title: Assessment of porphyrogenicity of drugs and chemicals in selected hepatic cell culture models through a fluorescence‐based screening assay. Assessment of porphyrogenicity of drugs and chemicals in selected hepatic cell culture models through a fluorescence‐based screening assay

    Journal: Pharmacology Research & Perspectives

    doi: 10.1002/prp2.951

    Fluorescence produced in various cell models after 18–24 h AIA or ALA treatment. Porphyrin accumulation produced by increasing concentrations of ALA, ranging from 0 to 1 mM or with 0.314 mM AIA, all in the presence and absence of 250‐μM DFO is displayed in (A) LMH cells and (B) HepG2 cells. (C) Fluorescence readings produced in the presence of 0.5‐mM AIA, ALA, phenobarbital, and phenytoin with and without DFO are shown in HepG2 spheroids. All fluorescence readings were corrected by deducting background readings. Data are presented as mean values ± SEM of three independent replicates. A two‐sided Student's t ‐test was used to assess the statistical significance between the two ratios
    Figure Legend Snippet: Fluorescence produced in various cell models after 18–24 h AIA or ALA treatment. Porphyrin accumulation produced by increasing concentrations of ALA, ranging from 0 to 1 mM or with 0.314 mM AIA, all in the presence and absence of 250‐μM DFO is displayed in (A) LMH cells and (B) HepG2 cells. (C) Fluorescence readings produced in the presence of 0.5‐mM AIA, ALA, phenobarbital, and phenytoin with and without DFO are shown in HepG2 spheroids. All fluorescence readings were corrected by deducting background readings. Data are presented as mean values ± SEM of three independent replicates. A two‐sided Student's t ‐test was used to assess the statistical significance between the two ratios

    Techniques Used: Fluorescence, Produced

    Porphyrogenicity of selected compounds in LMH cells after 18–24 h treatment. Fluorescence readings produced by the positive and negative controls, 0.314‐mM AIA and 0.314‐mM acetaminophen, respectively, represent clear levels of porphyrogenicity and non‐porphyrogenicity, respectively, in LMH cells. Among all the compounds, higher concentrations of ALA, norgestrel, phenobarbital, phenytoin, and sodium valproate produced fluorescence readings greater than the readings produced by AIA. Data are presented as mean values ± SEM of three independent replicates. All results are representative of three independent experiments. A two‐sided Student's t ‐test was used to assess the statistical significance between the two ratios
    Figure Legend Snippet: Porphyrogenicity of selected compounds in LMH cells after 18–24 h treatment. Fluorescence readings produced by the positive and negative controls, 0.314‐mM AIA and 0.314‐mM acetaminophen, respectively, represent clear levels of porphyrogenicity and non‐porphyrogenicity, respectively, in LMH cells. Among all the compounds, higher concentrations of ALA, norgestrel, phenobarbital, phenytoin, and sodium valproate produced fluorescence readings greater than the readings produced by AIA. Data are presented as mean values ± SEM of three independent replicates. All results are representative of three independent experiments. A two‐sided Student's t ‐test was used to assess the statistical significance between the two ratios

    Techniques Used: Fluorescence, Produced

    Cytotoxicity of compounds in LMH cells after 18–24 h treatment. A parallel plate with the same treatment as the fluorescence‐based in vitro drug screening assay was assayed for cytotoxicity with the ATPLite cytotoxicity assay. CC 50 values were calculated using the software, GraphPad Prism 8. Data are presented as mean values ± SEM of three independent replicates. All results are representative of three independent experiments
    Figure Legend Snippet: Cytotoxicity of compounds in LMH cells after 18–24 h treatment. A parallel plate with the same treatment as the fluorescence‐based in vitro drug screening assay was assayed for cytotoxicity with the ATPLite cytotoxicity assay. CC 50 values were calculated using the software, GraphPad Prism 8. Data are presented as mean values ± SEM of three independent replicates. All results are representative of three independent experiments

    Techniques Used: Fluorescence, In Vitro, Screening Assay, Cytotoxicity Assay, Software

    25) Product Images from "Systematic identification of chicken type I, II and III interferon-stimulated genes"

    Article Title: Systematic identification of chicken type I, II and III interferon-stimulated genes

    Journal: Veterinary Research

    doi: 10.1186/s13567-020-00793-x

    Validation of RNA-Seq data by qPCR. ISGs were selected from ChIFN-α-stimulated DF1 cells ( A ); ChIFN-γ- stimulated DF1 cells ( B ); ChIFN-λ- stimulated DF1 cells ( C ); and ChIFN-λ- stimulated LMH cells ( D ). The data of relative mRNA expression level was derived from the ratio of the ChIFN-treatment group results to the control group results. qPCR and RNA-seq results were respectively displayed as 2 −ΔΔCt value and the average log 2 (fold change) values of DEG. Data from qPCR are representative of three independent experiments. * p
    Figure Legend Snippet: Validation of RNA-Seq data by qPCR. ISGs were selected from ChIFN-α-stimulated DF1 cells ( A ); ChIFN-γ- stimulated DF1 cells ( B ); ChIFN-λ- stimulated DF1 cells ( C ); and ChIFN-λ- stimulated LMH cells ( D ). The data of relative mRNA expression level was derived from the ratio of the ChIFN-treatment group results to the control group results. qPCR and RNA-seq results were respectively displayed as 2 −ΔΔCt value and the average log 2 (fold change) values of DEG. Data from qPCR are representative of three independent experiments. * p

    Techniques Used: RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Expressing, Derivative Assay

    Systematic identification of chicken type I, II and III ISGs. A Differentially expressed genes (DEGs) in chicken IFN-stimulated cells as detected by RNA-Seq. B Venn diagrams of up-regulated DEGs in DF1 and LMH cells stimulated with ChIFN-λ. Up-regulated genes induced by ChIFN-λ in both DF1 and LMH cells were considered as chicken type III IFN-stimulated genes (ISGs). C Statistics used for the identification of chicken type I, II, and III ISGs in this study.
    Figure Legend Snippet: Systematic identification of chicken type I, II and III ISGs. A Differentially expressed genes (DEGs) in chicken IFN-stimulated cells as detected by RNA-Seq. B Venn diagrams of up-regulated DEGs in DF1 and LMH cells stimulated with ChIFN-λ. Up-regulated genes induced by ChIFN-λ in both DF1 and LMH cells were considered as chicken type III IFN-stimulated genes (ISGs). C Statistics used for the identification of chicken type I, II, and III ISGs in this study.

    Techniques Used: RNA Sequencing Assay

    Preparation of chicken types II and III interferon. A Chicken IFN-γ and IFN-λ (ChIFN-γ and ChIFN-λ) were prepared after expression in HEK293E cells. Purified ChIFN-γ and ChIFN-λ samples were subjected to SDS-PAGE. B The purified proteins were further verified by Western blotting using His antibody and ChIFN-γ and ChIFN-λ serum polyclonal antibody. C The bioactivity of the purified ChIFN-γ and ChIFN-λ proteins was detected in DF1 and LMH cell lines. The results are representative of three independent experiments.
    Figure Legend Snippet: Preparation of chicken types II and III interferon. A Chicken IFN-γ and IFN-λ (ChIFN-γ and ChIFN-λ) were prepared after expression in HEK293E cells. Purified ChIFN-γ and ChIFN-λ samples were subjected to SDS-PAGE. B The purified proteins were further verified by Western blotting using His antibody and ChIFN-γ and ChIFN-λ serum polyclonal antibody. C The bioactivity of the purified ChIFN-γ and ChIFN-λ proteins was detected in DF1 and LMH cell lines. The results are representative of three independent experiments.

    Techniques Used: Expressing, Purification, SDS Page, Western Blot

    Activity analysis of IFN-stimulated response element (ISRE) and gamma-activated sequence (GAS) elements after stimulation by chicken IFN. A – C Analysis of activation of ISRE and GAS elements induced by chicken type I, II, and III IFNs in DF1 cells. D Analysis of activation of ISRE and GAS elements induced by chicken type III IFN in LMH cells. Data are representative of three independent experiments. ns: not significant ( P > 0.05), * P
    Figure Legend Snippet: Activity analysis of IFN-stimulated response element (ISRE) and gamma-activated sequence (GAS) elements after stimulation by chicken IFN. A – C Analysis of activation of ISRE and GAS elements induced by chicken type I, II, and III IFNs in DF1 cells. D Analysis of activation of ISRE and GAS elements induced by chicken type III IFN in LMH cells. Data are representative of three independent experiments. ns: not significant ( P > 0.05), * P

    Techniques Used: Activity Assay, Sequencing, Activation Assay

    26) Product Images from "Requirement of Cellular Protein CCT7 for the Replication of Fowl Adenovirus Serotype 4 (FAdV-4) in Leghorn Male Hepatocellular Cells Via Interaction with the Viral Hexon Protein"

    Article Title: Requirement of Cellular Protein CCT7 for the Replication of Fowl Adenovirus Serotype 4 (FAdV-4) in Leghorn Male Hepatocellular Cells Via Interaction with the Viral Hexon Protein

    Journal: Viruses

    doi: 10.3390/v11020107

    Interaction of FAdV-4 hexon with cellular protein CCT7. ( A , B ) The interaction of hexon with exogenous CCT7. LMH cells ( A ) and HeLa cells ( B ) were transfected with the indicated expression plasmids and immunoprecipitated (IP) with anti-FLAG antibodies. Hexon and CCT7 in the immune complex were examined with Western Blot using anti-FLAG and anti-Myc McAbs, respectively in the cell lysates and in the immunoprecipitates too. ( C ) The interaction of hexon with endogenous CCT7 in LMH cells. LMH cells were transfected with pRK5-FLAG-hexon or empty vector as control. Twenty-four hours after transfection, cell lysates were prepared and immunoprecipitated with anti-FLAG antibody. Both the cell lysates and the immunoprecipitates were immunoblotted with anti-FLAG or anti-CCT7 antibodies. ( D ) Interaction of hexon with endogenous CCT7 in FAdV-4-infected cells. LMH cells were mock infected or infected with FAdV-4 at an MOI of 1, and immunoprecipitation assays were performed with an anti-hexon McAb. CCT7 in the immune complex was examined with Western Blot using anti-CCT7 polyclonal antibodies. The “H chain” means the heavy chain of the antibody. The “isotype” means isotype (IgG2b) control antibody of anti-FAdV-4 hexon monoclonal antibody.
    Figure Legend Snippet: Interaction of FAdV-4 hexon with cellular protein CCT7. ( A , B ) The interaction of hexon with exogenous CCT7. LMH cells ( A ) and HeLa cells ( B ) were transfected with the indicated expression plasmids and immunoprecipitated (IP) with anti-FLAG antibodies. Hexon and CCT7 in the immune complex were examined with Western Blot using anti-FLAG and anti-Myc McAbs, respectively in the cell lysates and in the immunoprecipitates too. ( C ) The interaction of hexon with endogenous CCT7 in LMH cells. LMH cells were transfected with pRK5-FLAG-hexon or empty vector as control. Twenty-four hours after transfection, cell lysates were prepared and immunoprecipitated with anti-FLAG antibody. Both the cell lysates and the immunoprecipitates were immunoblotted with anti-FLAG or anti-CCT7 antibodies. ( D ) Interaction of hexon with endogenous CCT7 in FAdV-4-infected cells. LMH cells were mock infected or infected with FAdV-4 at an MOI of 1, and immunoprecipitation assays were performed with an anti-hexon McAb. CCT7 in the immune complex was examined with Western Blot using anti-CCT7 polyclonal antibodies. The “H chain” means the heavy chain of the antibody. The “isotype” means isotype (IgG2b) control antibody of anti-FAdV-4 hexon monoclonal antibody.

    Techniques Used: Transfection, Expressing, Immunoprecipitation, Western Blot, Plasmid Preparation, Infection

    The portion of CCT7 from amino acids 137–272 is responsible for binding to hexon. ( A ) Schematics represent the genes encoding the full-length CCT7 and truncated CCT7 molecules (from Δ1 to Δ6). ( B ) Interaction of Flag-hexon with different truncated CCT7 mutants. LMH cells were transfected with full-length pCMV-myc-cct7, the indicated truncated mutants or pCMV-myc-control (encode a 17kDa control protein). Cell lysates were prepared and immunoprecipitated with anti-Flag McAb. The pellets were examined with Western Blot using anti-Myc and anti-FLAG antibodies. ( C ) Schematic representing the domain (137–272aa) of CCT7 responsible for binding to hexon. The “H chain” means the heavy chain of antibody and the “L chain” means the light chain of antibody.
    Figure Legend Snippet: The portion of CCT7 from amino acids 137–272 is responsible for binding to hexon. ( A ) Schematics represent the genes encoding the full-length CCT7 and truncated CCT7 molecules (from Δ1 to Δ6). ( B ) Interaction of Flag-hexon with different truncated CCT7 mutants. LMH cells were transfected with full-length pCMV-myc-cct7, the indicated truncated mutants or pCMV-myc-control (encode a 17kDa control protein). Cell lysates were prepared and immunoprecipitated with anti-Flag McAb. The pellets were examined with Western Blot using anti-Myc and anti-FLAG antibodies. ( C ) Schematic representing the domain (137–272aa) of CCT7 responsible for binding to hexon. The “H chain” means the heavy chain of antibody and the “L chain” means the light chain of antibody.

    Techniques Used: Binding Assay, Transfection, Immunoprecipitation, Western Blot

    Screening for hexon-interacting proteins in LMH cells. ( A – C ) LMH cells transfected with pRK5-flag-hexon or pRK5-flag and subjected to a pull-down assay ( A ). The arrow-pointed protein bands in ( A ) were further analyzed by liquid chromatography-mass spectrograph (LC-MS), and the cellular protein CCT7 was identified from the arrow-pointed protein band c ( B ). The alignment of the chicken CCT7 sequences (UniProtKB: A0A1L1RLC2) with its matched peptides is shown in bold red as identified by LC-MS ( C ).
    Figure Legend Snippet: Screening for hexon-interacting proteins in LMH cells. ( A – C ) LMH cells transfected with pRK5-flag-hexon or pRK5-flag and subjected to a pull-down assay ( A ). The arrow-pointed protein bands in ( A ) were further analyzed by liquid chromatography-mass spectrograph (LC-MS), and the cellular protein CCT7 was identified from the arrow-pointed protein band c ( B ). The alignment of the chicken CCT7 sequences (UniProtKB: A0A1L1RLC2) with its matched peptides is shown in bold red as identified by LC-MS ( C ).

    Techniques Used: Transfection, Pull Down Assay, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy

    CCT7 is required for the presence of hexon. ( A , B ) Overexpression of CCT7 enhanced hexon levels in cells transfected with pRK5-flag-hexon. LMH cells were transfected with the indicated plasmids; twenty-four hours after transfection, the cell lysates were prepared and examined by Western Blot ( A ). The levels of hexon in ( A ) were quantified by densitometry and normalized to that of GAPDH ( B ). The level of hexon in controls was set at 1.0. ( C , D ) Effects of CCT7 RNAi on the expression of endogenous CCT7. LMH cells were transfected with siRNA (RNAi#1-3) or controls. Twenty-four hours after the second transfection, cell lysates were prepared and examined by Western Blot using anti-CCT7 antibodies ( C ). Endogenous GAPDH was used as an internal control. The density of bands in ( C ) was quantitated by densitometry as above described ( D ). ( E , F ) knockdown of CCT7 reduced hexon expression in cells. LMH cells were treated with CCT7 RNAi or control RNAi and transfected with pRK5-flag-hexon ( E ). Twenty-four hours post-transfection, cell lysates were prepared, and the expression levels of hexon were examined by Western Blot using anti-hexon McAb. The levels of hexon expression in ( E ) were quantified by densitometry and normalized to that of GAPDH ( F ). The expression levels of hexon in normal control LMH cells was set at 1.0. ( G , H ) LMH cells were treated with CCT7 RNAi or control RNAi, followed by infection with FAdV-4 at an MOI of 0.1. Twenty-four hours post-infection, cell lysates were prepared, and the expression of hexon was examined by Western Blot using anti-hexon McAb ( G ). The levels of hexon expression in ( G ) were quantified by densitometry and normalized to that of GAPDH ( H ). The expression level of hexon in normal control LMH cells was set at 1.0. Data are represented as mean ± SD, n = 3. ** stands for p
    Figure Legend Snippet: CCT7 is required for the presence of hexon. ( A , B ) Overexpression of CCT7 enhanced hexon levels in cells transfected with pRK5-flag-hexon. LMH cells were transfected with the indicated plasmids; twenty-four hours after transfection, the cell lysates were prepared and examined by Western Blot ( A ). The levels of hexon in ( A ) were quantified by densitometry and normalized to that of GAPDH ( B ). The level of hexon in controls was set at 1.0. ( C , D ) Effects of CCT7 RNAi on the expression of endogenous CCT7. LMH cells were transfected with siRNA (RNAi#1-3) or controls. Twenty-four hours after the second transfection, cell lysates were prepared and examined by Western Blot using anti-CCT7 antibodies ( C ). Endogenous GAPDH was used as an internal control. The density of bands in ( C ) was quantitated by densitometry as above described ( D ). ( E , F ) knockdown of CCT7 reduced hexon expression in cells. LMH cells were treated with CCT7 RNAi or control RNAi and transfected with pRK5-flag-hexon ( E ). Twenty-four hours post-transfection, cell lysates were prepared, and the expression levels of hexon were examined by Western Blot using anti-hexon McAb. The levels of hexon expression in ( E ) were quantified by densitometry and normalized to that of GAPDH ( F ). The expression levels of hexon in normal control LMH cells was set at 1.0. ( G , H ) LMH cells were treated with CCT7 RNAi or control RNAi, followed by infection with FAdV-4 at an MOI of 0.1. Twenty-four hours post-infection, cell lysates were prepared, and the expression of hexon was examined by Western Blot using anti-hexon McAb ( G ). The levels of hexon expression in ( G ) were quantified by densitometry and normalized to that of GAPDH ( H ). The expression level of hexon in normal control LMH cells was set at 1.0. Data are represented as mean ± SD, n = 3. ** stands for p

    Techniques Used: Over Expression, Transfection, Western Blot, Expressing, Infection

    Effects of CCT7 on FAdV-4 growth in LMH cells. ( A ) Knockdown of CCT7 suppresses FAdV-4 growth. LMH cells were treated with RNAi to knockdown endogenous CCT7 expression as described above, followed by infection with FAdV-4 at an MOI of 1. At different time points (12, 24, 48, and 72 h) post FAdV-4 infection, the viral titers in the cell cultures were determined by TCID50 analysis using 96-well plates. The significance of the difference between CCT7 RNAi treatment and RNAi controls was performed by ANOVA. ( B ) LMH cells were transfected with pCMV-myc-cct7 or empty vectors, followed by infection with FAdV-4 at an MOI of 1. At different time points (12, 24, 48, and 72 h) post FAdV-4 infection, the viral loads in the cell cultures were determined by TCID 50 using 96-well plates. The significance of the difference between pCMV-myc-cct7 transfected cells and controls in viral growth was determined by ANOVA. * stands for p
    Figure Legend Snippet: Effects of CCT7 on FAdV-4 growth in LMH cells. ( A ) Knockdown of CCT7 suppresses FAdV-4 growth. LMH cells were treated with RNAi to knockdown endogenous CCT7 expression as described above, followed by infection with FAdV-4 at an MOI of 1. At different time points (12, 24, 48, and 72 h) post FAdV-4 infection, the viral titers in the cell cultures were determined by TCID50 analysis using 96-well plates. The significance of the difference between CCT7 RNAi treatment and RNAi controls was performed by ANOVA. ( B ) LMH cells were transfected with pCMV-myc-cct7 or empty vectors, followed by infection with FAdV-4 at an MOI of 1. At different time points (12, 24, 48, and 72 h) post FAdV-4 infection, the viral loads in the cell cultures were determined by TCID 50 using 96-well plates. The significance of the difference between pCMV-myc-cct7 transfected cells and controls in viral growth was determined by ANOVA. * stands for p

    Techniques Used: Expressing, Infection, Transfection

    Involvement of CCT7 in hexon reduction in LMH cells. ( A , B ) LMH cells were cotransfected with pCMV-Myc-cct7 and pRK5-flag-hexon or control empty vector. Twelve hours after transfection, cells were treated with cycloheximide (CHX, 50 μg/mL) at different time points (0, 1, 2, 3, 4, and 5 h) before being harvested. Cell lysates were prepared and examined with Western Blot using the indicated antibodies. The levels of Flag-hexon in ( A ) were quantified by densitometry and normalized to that of GAPDH ( B ). The value of normalized Flag-hexon at time 0 h was set at 1.0. ( C , D ) LMH cells were treated with CCT7 RNAi or control RNAi and transfected with pRK5-flag-hexon. Twelve hours after transfection, cells were treated with cycloheximide (CHX, 50 μg/mL) at different time points (0, 0.5, 1.0, 1.5, 2.5, and 3.5 h) before being harvested. Cell lysates were prepared and examined with Western Blot using indicated antibodies. The levels of Flag-hexon in ( C ) were quantified by densitometry and normalized to that of GAPDH ( D ). The value of normalized Flag-hexon at time point 0 h was set at 1.0. The significance of the difference was determined by ANOVA.
    Figure Legend Snippet: Involvement of CCT7 in hexon reduction in LMH cells. ( A , B ) LMH cells were cotransfected with pCMV-Myc-cct7 and pRK5-flag-hexon or control empty vector. Twelve hours after transfection, cells were treated with cycloheximide (CHX, 50 μg/mL) at different time points (0, 1, 2, 3, 4, and 5 h) before being harvested. Cell lysates were prepared and examined with Western Blot using the indicated antibodies. The levels of Flag-hexon in ( A ) were quantified by densitometry and normalized to that of GAPDH ( B ). The value of normalized Flag-hexon at time 0 h was set at 1.0. ( C , D ) LMH cells were treated with CCT7 RNAi or control RNAi and transfected with pRK5-flag-hexon. Twelve hours after transfection, cells were treated with cycloheximide (CHX, 50 μg/mL) at different time points (0, 0.5, 1.0, 1.5, 2.5, and 3.5 h) before being harvested. Cell lysates were prepared and examined with Western Blot using indicated antibodies. The levels of Flag-hexon in ( C ) were quantified by densitometry and normalized to that of GAPDH ( D ). The value of normalized Flag-hexon at time point 0 h was set at 1.0. The significance of the difference was determined by ANOVA.

    Techniques Used: Plasmid Preparation, Transfection, Western Blot

    Replication of FAdV-4 strain HuBWH in LMH cells. ( A – D ) LMH cells were mock infected (A and C) or infected with FAdV-4 at an MOI of 1 ( B , D ). Twenty-four hours after FAdV-4 infection, cells were subjected to IFA staining using mouse anti-hexon polyclonal antibodies followed by FITC-conjugated goat anti-mouse IgG antibodies and were visualized under a fluorescence microscope. Magnification, ×400. ( E ) Detection of the viral protein hexon in FAdV-4-infected or mock-infected LMH cells by Western Blot using an anti-hexon McAb. ( F ) Examination of FAdV-4 replication in LMH cells. LMH cells were infected with FAdV-4 at an MOI of 1. At different time points (12, 24, 48, and 72 h) after FAdV-4 infection, the viral titers in the cell cultures were determined by TCID 50 . (G to I) Examination of FAdV-4 HuBWH particles by TEM. LMH cells were mock infected ( G ) or infected with FAdV-4 at an MOI of 1 for 24 h ( H ). A higher-magnification view of panel H is demonstrated by ( I ) (The FAdV-4 particles are indicated by red arrows). Scale bar = 1 μm in G H; Scale bar = 500 nm in I.
    Figure Legend Snippet: Replication of FAdV-4 strain HuBWH in LMH cells. ( A – D ) LMH cells were mock infected (A and C) or infected with FAdV-4 at an MOI of 1 ( B , D ). Twenty-four hours after FAdV-4 infection, cells were subjected to IFA staining using mouse anti-hexon polyclonal antibodies followed by FITC-conjugated goat anti-mouse IgG antibodies and were visualized under a fluorescence microscope. Magnification, ×400. ( E ) Detection of the viral protein hexon in FAdV-4-infected or mock-infected LMH cells by Western Blot using an anti-hexon McAb. ( F ) Examination of FAdV-4 replication in LMH cells. LMH cells were infected with FAdV-4 at an MOI of 1. At different time points (12, 24, 48, and 72 h) after FAdV-4 infection, the viral titers in the cell cultures were determined by TCID 50 . (G to I) Examination of FAdV-4 HuBWH particles by TEM. LMH cells were mock infected ( G ) or infected with FAdV-4 at an MOI of 1 for 24 h ( H ). A higher-magnification view of panel H is demonstrated by ( I ) (The FAdV-4 particles are indicated by red arrows). Scale bar = 1 μm in G H; Scale bar = 500 nm in I.

    Techniques Used: Infection, Immunofluorescence, Staining, Fluorescence, Microscopy, Western Blot, Transmission Electron Microscopy

    Colocalization of hexon with CCT7 in LMH cells. ( A – E ) Localization of hexon and exogenous CCT7 in LMH cells. LMH cells were seeded onto 24-well plates with coverslips in the wells and cultured overnight. Cells were transfected with pEGFP-hexon ( A ), pDsRed-cct7 ( b ) or both pEGFP-hexon and pDsRed-cct7 ( C – E ). Twenty-four hours after infection, cells were fixed and the cell nuclei were counterstained with DAPI (blue). The cell samples were observed with a confocal laser scanning microscope. ( F – K ) Colocalization of FAdV-4 hexon with exogenous CCT7 in FAdV-4-infected cells. LMH cells were mock infected or infected with FAdV-4 at an MOI of 1. Two hours after infection, cells were transfected with the indicated expression plasmids. Twenty-two hours after transfection, cells were fixed and probed with mouse anti-hexon polyclonal antibodies, followed by incubation with FITC-conjugated goat anti-mouse antibodies (green). Nuclei were counterstained with DAPI (blue). ( L – Q ) Colocalization of FAdV-4 hexon with endogenous CCT7 in FAdV-4-infected cells. LMH cells were mock infected or infected with FAdV-4 at an MOI of 1. Twenty-four hours after infection, cells were fixed and probed with mouse anti-hexon and rabbit anti-CCT7 polyclonal antibodies, followed by incubation with TRITC-conjugated goat anti-mouse antibodies (red) and DyLight 488 affiniPure goat anti-rabbit IgG antibodies (green). Nuclei were counterstained with DAPI (blue). The cell samples were observed with a confocal laser scanning microscope. Scale bar = 5 μm.
    Figure Legend Snippet: Colocalization of hexon with CCT7 in LMH cells. ( A – E ) Localization of hexon and exogenous CCT7 in LMH cells. LMH cells were seeded onto 24-well plates with coverslips in the wells and cultured overnight. Cells were transfected with pEGFP-hexon ( A ), pDsRed-cct7 ( b ) or both pEGFP-hexon and pDsRed-cct7 ( C – E ). Twenty-four hours after infection, cells were fixed and the cell nuclei were counterstained with DAPI (blue). The cell samples were observed with a confocal laser scanning microscope. ( F – K ) Colocalization of FAdV-4 hexon with exogenous CCT7 in FAdV-4-infected cells. LMH cells were mock infected or infected with FAdV-4 at an MOI of 1. Two hours after infection, cells were transfected with the indicated expression plasmids. Twenty-two hours after transfection, cells were fixed and probed with mouse anti-hexon polyclonal antibodies, followed by incubation with FITC-conjugated goat anti-mouse antibodies (green). Nuclei were counterstained with DAPI (blue). ( L – Q ) Colocalization of FAdV-4 hexon with endogenous CCT7 in FAdV-4-infected cells. LMH cells were mock infected or infected with FAdV-4 at an MOI of 1. Twenty-four hours after infection, cells were fixed and probed with mouse anti-hexon and rabbit anti-CCT7 polyclonal antibodies, followed by incubation with TRITC-conjugated goat anti-mouse antibodies (red) and DyLight 488 affiniPure goat anti-rabbit IgG antibodies (green). Nuclei were counterstained with DAPI (blue). The cell samples were observed with a confocal laser scanning microscope. Scale bar = 5 μm.

    Techniques Used: Cell Culture, Transfection, Infection, Laser-Scanning Microscopy, Expressing, Incubation

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    ATCC lmh cells
    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. <t>LMH</t> <t>cells</t> transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.
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    Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Journal: Frontiers in Veterinary Science

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    doi: 10.3389/fvets.2022.1003262

    Figure Lengend Snippet: Mapping the epitopes in Fiber-1 protein recognized by these mAbs. LMH cells transfected with the different Fiber-1 truncations were tested using mAb 6F10, 3G6, 4G1, 1D8, 4G2, and 1E6 by IFA.

    Article Snippet: These recombinant plasmids were expressed in LMH cells and the epitopes recognized by the mAbs were identified using IFA.

    Techniques: Transfection, Immunofluorescence

    Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Journal: Frontiers in Veterinary Science

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    doi: 10.3389/fvets.2022.1003262

    Figure Lengend Snippet: Characterization analysis of the generated mAbs by IFA and Western blot. (A) The mAbs reacted with LMH cells infected with DAdV-3, but not with mock-infected LMH cells (NC) by IFA. (B) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1, but not with LMH cells transfected with pcDNA3.1 (NC) by IFA. (C) The mAbs reacted with LMH cells infected with DAdV-3 by Western blot. Lane 1 and 2: LMH cells infected without or with DAdV-3, respectively. (D) The mAbs reacted with LMH cells transfected with pcDNA3.1-Fiber-1 by Western blot. Lane 1 and 2: LMH cells transfected with pcDNA3.1 or pcDNA3.1-Fiber-1, respectively.

    Article Snippet: These recombinant plasmids were expressed in LMH cells and the epitopes recognized by the mAbs were identified using IFA.

    Techniques: Generated, Immunofluorescence, Western Blot, Infection, Transfection

    mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Journal: Frontiers in Veterinary Science

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    doi: 10.3389/fvets.2022.1003262

    Figure Lengend Snippet: mAbs 3G6 and 6F10 efficiently immunoprecipitated Fiber-1 of DAdV-3. (A) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells transfected with pcDNA3.1-Fiber-1. Lane 1 and 4: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 and pcDNA3.1-Fiber-1, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells transfected with pcDNA3.1-Fiber-1 immunoprecipitated by mAb 3G6 and 6F10, respectively. (B) mAbs 3G6 and 6F10 immunoprecipitated Fiber-1 in LMH cells infected with DAdV-3. Lane 1 and 4: Western blot assay for the lysates of LMH cells infected without and with DAdV-3, respectively. Lanes 2 and 3: Western blot assay for the lysates of LMH cells without the infection of DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively. Lanes 5 and 6: Western blot assay for the lysates of LMH cells infected with DAdV-3 immunoprecipitated by mAb 3G6 and 6F10, respectively.

    Article Snippet: These recombinant plasmids were expressed in LMH cells and the epitopes recognized by the mAbs were identified using IFA.

    Techniques: Immunoprecipitation, Transfection, Western Blot, Infection

    Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Journal: Frontiers in Veterinary Science

    Article Title: Novel monoclonal antibodies against Fiber-1 of duck adenovirus 3 and their B cell epitopes

    doi: 10.3389/fvets.2022.1003262

    Figure Lengend Snippet: Neutralization activities of mAbs 3G6 and 4G2 against DAdV-3. (A) Western blot assay for the neutralizing activity of mAbs 3G6 and 4G2. Lanes 1 and 2: The lysates of LMH cells infected without or with DAdV-3. Lane 3 and 4: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 3G6, respectively. Lane 5 and 6: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of mAb 4G2, respectively. Lane 7 and 8: The lysates of the infected LMH cells treated with 10 and 20 μg/ml of the control mAb 3G12, respectively. (B) Analysis of the rate of the expressed level of Fiber-1 and GAPDH in the neutralizing test.

    Article Snippet: These recombinant plasmids were expressed in LMH cells and the epitopes recognized by the mAbs were identified using IFA.

    Techniques: Neutralization, Western Blot, Activity Assay, Infection