Structured Review

Funakoshi streptavidin
Detection of the intracellular DPE-modified proteins. (A) Detection of the DPE-modified proteins in Hepa1c1c7. Confluent Hepa1c1c7 cells were pre-incubated with 0.1% DMSO or 100 µM BNPP and incubated with 100 µM DPE for indicated time periods in serum-free MEM-α. The DPE-tagged cellular proteins were detected by <t>HRP-streptavidin.</t> (B) Structural comparison of DBE with DOPAC and DPE. (C) Effect of the pre-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in Hepa1c1c7 cells. Hepa1c1c7 cells were pre-incubated with DOPAC (open circle) or DBE (closed circle) for 30 min and incubated with 25 µM DPE for 3 h in serum-free MEM-α. (D) Effect of the co-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in cell lysate. Cell lysate was incubated with 25 µM DPE in combination with DOPAC (open circle) or DBE (closed circle) in the presence of 30 U laccase for 1 h. The values represent means ±S.D. of three separate experiments.
Streptavidin, supplied by Funakoshi, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 1 article reviews
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streptavidin - by Bioz Stars, 2020-04
92/100 stars

Images

1) Product Images from "A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite"

Article Title: A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite

Journal: Biochemistry and Biophysics Reports

doi: 10.1016/j.bbrep.2016.06.020

Detection of the intracellular DPE-modified proteins. (A) Detection of the DPE-modified proteins in Hepa1c1c7. Confluent Hepa1c1c7 cells were pre-incubated with 0.1% DMSO or 100 µM BNPP and incubated with 100 µM DPE for indicated time periods in serum-free MEM-α. The DPE-tagged cellular proteins were detected by HRP-streptavidin. (B) Structural comparison of DBE with DOPAC and DPE. (C) Effect of the pre-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in Hepa1c1c7 cells. Hepa1c1c7 cells were pre-incubated with DOPAC (open circle) or DBE (closed circle) for 30 min and incubated with 25 µM DPE for 3 h in serum-free MEM-α. (D) Effect of the co-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in cell lysate. Cell lysate was incubated with 25 µM DPE in combination with DOPAC (open circle) or DBE (closed circle) in the presence of 30 U laccase for 1 h. The values represent means ±S.D. of three separate experiments.
Figure Legend Snippet: Detection of the intracellular DPE-modified proteins. (A) Detection of the DPE-modified proteins in Hepa1c1c7. Confluent Hepa1c1c7 cells were pre-incubated with 0.1% DMSO or 100 µM BNPP and incubated with 100 µM DPE for indicated time periods in serum-free MEM-α. The DPE-tagged cellular proteins were detected by HRP-streptavidin. (B) Structural comparison of DBE with DOPAC and DPE. (C) Effect of the pre-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in Hepa1c1c7 cells. Hepa1c1c7 cells were pre-incubated with DOPAC (open circle) or DBE (closed circle) for 30 min and incubated with 25 µM DPE for 3 h in serum-free MEM-α. (D) Effect of the co-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in cell lysate. Cell lysate was incubated with 25 µM DPE in combination with DOPAC (open circle) or DBE (closed circle) in the presence of 30 U laccase for 1 h. The values represent means ±S.D. of three separate experiments.

Techniques Used: Modification, Incubation

CuAAC reaction of DPE with an azide-linked tag molecule. (A) Formation of 1,2,3-triazole by CuAAC reaction with DPE and benzyl azide. (B) Detection of GAPDH tagged by CuAAC reaction with DPE and the azide-labeled biotin. GAPDH (1 mg/ml) was incubated with or without 50 µM DPE in the presence or absence of 30 U laccase in 70 mM sodium phosphate buffer (pH 7.2) for 1 h at 37 °C. DPE-tagged GAPDH was detected by CBB staining (left) and HRP-streptavidin (right).
Figure Legend Snippet: CuAAC reaction of DPE with an azide-linked tag molecule. (A) Formation of 1,2,3-triazole by CuAAC reaction with DPE and benzyl azide. (B) Detection of GAPDH tagged by CuAAC reaction with DPE and the azide-labeled biotin. GAPDH (1 mg/ml) was incubated with or without 50 µM DPE in the presence or absence of 30 U laccase in 70 mM sodium phosphate buffer (pH 7.2) for 1 h at 37 °C. DPE-tagged GAPDH was detected by CBB staining (left) and HRP-streptavidin (right).

Techniques Used: Labeling, Incubation, Staining

Identification of the target proteins of DOPAC. Detection of the DPE-modified Keap1 and AhR. Confluent Hepa1c1c7 cells were incubated with 50 µM DPE for 5 h in serum-free MEM-α. The cell lysate was incubated with Streptavidin Mag Sepharose beads for 30 min. The DPE-modified Keap1 (left) or AhR (right) were detected by immunoblot analysis.
Figure Legend Snippet: Identification of the target proteins of DOPAC. Detection of the DPE-modified Keap1 and AhR. Confluent Hepa1c1c7 cells were incubated with 50 µM DPE for 5 h in serum-free MEM-α. The cell lysate was incubated with Streptavidin Mag Sepharose beads for 30 min. The DPE-modified Keap1 (left) or AhR (right) were detected by immunoblot analysis.

Techniques Used: Modification, Incubation

2) Product Images from "Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies"

Article Title: Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies

Journal: Scientific Reports

doi: 10.1038/s41598-017-03208-8

Reactivity of mAbs against apoA-I in the lipid-free state or on reconstituted discoidal HDL by ELISA. Biotinylated Cys-apoA-I in the lipid-free state or on discoidal HDL were added to 96-well microplates coated with streptavidin. After washing, Ab#19-17, Ab#20-7, Ab#124-2, or Ab#187-2 were added to the plates with increasing concentrations, and then detected with POD-conjugated anti-mouse IgG antibody.
Figure Legend Snippet: Reactivity of mAbs against apoA-I in the lipid-free state or on reconstituted discoidal HDL by ELISA. Biotinylated Cys-apoA-I in the lipid-free state or on discoidal HDL were added to 96-well microplates coated with streptavidin. After washing, Ab#19-17, Ab#20-7, Ab#124-2, or Ab#187-2 were added to the plates with increasing concentrations, and then detected with POD-conjugated anti-mouse IgG antibody.

Techniques Used: Enzyme-linked Immunosorbent Assay

Sandwich ELISA for detection of lipid-free and HDL-bound apoA-I. ApoA-I in the lipid-free state ( A ), on reconstituted discoidal HDL ( B ) and spherical HDL ( C ), and human plasma HDL ( D ) were added to the plates coated with Ab#124-2. After washing, biotinylated Ab#19-17, Ab#20-7, or Ab#187-2 were added to the plates, and then detected with POD-conjugated streptavidin.
Figure Legend Snippet: Sandwich ELISA for detection of lipid-free and HDL-bound apoA-I. ApoA-I in the lipid-free state ( A ), on reconstituted discoidal HDL ( B ) and spherical HDL ( C ), and human plasma HDL ( D ) were added to the plates coated with Ab#124-2. After washing, biotinylated Ab#19-17, Ab#20-7, or Ab#187-2 were added to the plates, and then detected with POD-conjugated streptavidin.

Techniques Used: Sandwich ELISA

Reactivity of the present monoclonal antibodies to full-length and the N-terminal 1–83 fragment of apoA-I. Biotinylated full-length apoA-I or its N-terminal 1–83 fragment (0.3 μM) in G-PBS, or PBS (for examining the background value) was added to 96-well microplates coated with streptavidin, and then incubated for 30 min at 37 °C. After washing the wells with T-PBS, hybridoma supernatants, 500-fold diluted with G-PBS, were added to the wells (100 μl/well) and incubated at 37 °C for 30 min. After washing, POD-conjugated anti-mouse IgG antibody diluted at 1:5000 in G-PBS (100 μl/well) was added and incubated for 30 min at 37 °C. The wells were washed and bound POD activity was determined colorimetrically (see Materials and Methods). *** P
Figure Legend Snippet: Reactivity of the present monoclonal antibodies to full-length and the N-terminal 1–83 fragment of apoA-I. Biotinylated full-length apoA-I or its N-terminal 1–83 fragment (0.3 μM) in G-PBS, or PBS (for examining the background value) was added to 96-well microplates coated with streptavidin, and then incubated for 30 min at 37 °C. After washing the wells with T-PBS, hybridoma supernatants, 500-fold diluted with G-PBS, were added to the wells (100 μl/well) and incubated at 37 °C for 30 min. After washing, POD-conjugated anti-mouse IgG antibody diluted at 1:5000 in G-PBS (100 μl/well) was added and incubated for 30 min at 37 °C. The wells were washed and bound POD activity was determined colorimetrically (see Materials and Methods). *** P

Techniques Used: Incubation, Activity Assay

Related Articles

Enzyme-linked Immunosorbent Assay:

Article Title: Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies
Article Snippet: .. Single-antibody ELISA Microwells of the Costar microplates were coated overnight at 4 °C with 25 μg/ml streptavidin (Funakoshi, Tokyo, Japan) in PBS (100 μl/well), and then blocked by incubating with 1% Block Ace (DS Pharma Biomedical, Osaka, Japan) for 60 min at 37 °C. .. The wells were washed with T-PBS and then incubated with biotinylated apoA-I samples diluted with G-PBS (100 μl/well) for 30 min at 37 °C.

Protease Inhibitor:

Article Title: A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite
Article Snippet: 2.1 Materials DOPAC, laccase from Rhus vernicifera , glyceraldehyde-3-phosphate dehydrogenase (GAPDH), tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA), bis(4-nitrophenyl) phosphate (BNPP) and azide-PEG3 -biotin conjugate were obtained from Sigma Aldrich (St. Louis, USA). p -Toluensulfonic acid monohydrate (PTSA), n -butanol, toluene, copper (II) sulfate pentahydrate, protease inhibitor cocktail and Chemi-Lumi One Super were purchased from nacalai tasque (Kyoto, Japan). .. Streptavidin, HRP conjugate was purchased from Funakoshi (Tokyo, Japan).

Incubation:

Article Title: Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies
Article Snippet: Single-antibody ELISA Microwells of the Costar microplates were coated overnight at 4 °C with 25 μg/ml streptavidin (Funakoshi, Tokyo, Japan) in PBS (100 μl/well), and then blocked by incubating with 1% Block Ace (DS Pharma Biomedical, Osaka, Japan) for 60 min at 37 °C. .. The wells were washed with T-PBS and then incubated with biotinylated apoA-I samples diluted with G-PBS (100 μl/well) for 30 min at 37 °C.

Blocking Assay:

Article Title: Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies
Article Snippet: .. Single-antibody ELISA Microwells of the Costar microplates were coated overnight at 4 °C with 25 μg/ml streptavidin (Funakoshi, Tokyo, Japan) in PBS (100 μl/well), and then blocked by incubating with 1% Block Ace (DS Pharma Biomedical, Osaka, Japan) for 60 min at 37 °C. .. The wells were washed with T-PBS and then incubated with biotinylated apoA-I samples diluted with G-PBS (100 μl/well) for 30 min at 37 °C.

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    Funakoshi streptavidin conjugated horseradish peroxidase
    Immunoreactivity of the Ki-67 antibody in the nuclei of lung squamous cell carcinoma cells. (A) Low-grade cell proliferation, hematoxylin and eosin; (B) low-grade cell proliferation, LSAB method; (C) high-grade cell proliferation (≥30% Ki-67 LI), hematoxylin and eosin; (D) high-grade cell proliferation (≥30% Ki-67 LI), LSAB method. X25. LI, labeling index; LSAB, labeled <t>streptavidin-biotin.</t>
    Streptavidin Conjugated Horseradish Peroxidase, supplied by Funakoshi, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin conjugated horseradish peroxidase/product/Funakoshi
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin conjugated horseradish peroxidase - by Bioz Stars, 2020-04
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    92
    Funakoshi streptavidin
    Detection of the intracellular DPE-modified proteins. (A) Detection of the DPE-modified proteins in Hepa1c1c7. Confluent Hepa1c1c7 cells were pre-incubated with 0.1% DMSO or 100 µM BNPP and incubated with 100 µM DPE for indicated time periods in serum-free MEM-α. The DPE-tagged cellular proteins were detected by <t>HRP-streptavidin.</t> (B) Structural comparison of DBE with DOPAC and DPE. (C) Effect of the pre-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in Hepa1c1c7 cells. Hepa1c1c7 cells were pre-incubated with DOPAC (open circle) or DBE (closed circle) for 30 min and incubated with 25 µM DPE for 3 h in serum-free MEM-α. (D) Effect of the co-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in cell lysate. Cell lysate was incubated with 25 µM DPE in combination with DOPAC (open circle) or DBE (closed circle) in the presence of 30 U laccase for 1 h. The values represent means ±S.D. of three separate experiments.
    Streptavidin, supplied by Funakoshi, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin/product/Funakoshi
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    94
    Funakoshi horseradish peroxidase hrp conjugated streptavidin solution
    Detection of the intracellular DPE-modified proteins. (A) Detection of the DPE-modified proteins in Hepa1c1c7. Confluent Hepa1c1c7 cells were pre-incubated with 0.1% DMSO or 100 µM BNPP and incubated with 100 µM DPE for indicated time periods in serum-free MEM-α. The DPE-tagged cellular proteins were detected by <t>HRP-streptavidin.</t> (B) Structural comparison of DBE with DOPAC and DPE. (C) Effect of the pre-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in Hepa1c1c7 cells. Hepa1c1c7 cells were pre-incubated with DOPAC (open circle) or DBE (closed circle) for 30 min and incubated with 25 µM DPE for 3 h in serum-free MEM-α. (D) Effect of the co-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in cell lysate. Cell lysate was incubated with 25 µM DPE in combination with DOPAC (open circle) or DBE (closed circle) in the presence of 30 U laccase for 1 h. The values represent means ±S.D. of three separate experiments.
    Horseradish Peroxidase Hrp Conjugated Streptavidin Solution, supplied by Funakoshi, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/horseradish peroxidase hrp conjugated streptavidin solution/product/Funakoshi
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    horseradish peroxidase hrp conjugated streptavidin solution - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    Image Search Results


    Immunoreactivity of the Ki-67 antibody in the nuclei of lung squamous cell carcinoma cells. (A) Low-grade cell proliferation, hematoxylin and eosin; (B) low-grade cell proliferation, LSAB method; (C) high-grade cell proliferation (≥30% Ki-67 LI), hematoxylin and eosin; (D) high-grade cell proliferation (≥30% Ki-67 LI), LSAB method. X25. LI, labeling index; LSAB, labeled streptavidin-biotin.

    Journal: Molecular Medicine Reports

    Article Title: Ki-67 labeling index affects tumor infiltration patterns of lung squamous cell carcinoma

    doi: 10.3892/mmr.2015.4354

    Figure Lengend Snippet: Immunoreactivity of the Ki-67 antibody in the nuclei of lung squamous cell carcinoma cells. (A) Low-grade cell proliferation, hematoxylin and eosin; (B) low-grade cell proliferation, LSAB method; (C) high-grade cell proliferation (≥30% Ki-67 LI), hematoxylin and eosin; (D) high-grade cell proliferation (≥30% Ki-67 LI), LSAB method. X25. LI, labeling index; LSAB, labeled streptavidin-biotin.

    Article Snippet: The sections were then treated with streptavidin-conjugated horseradish peroxidase for 30 min at room temperature (Funakoshi Co., Ltd., Tokyo, Japan).

    Techniques: Labeling

    Detection of the intracellular DPE-modified proteins. (A) Detection of the DPE-modified proteins in Hepa1c1c7. Confluent Hepa1c1c7 cells were pre-incubated with 0.1% DMSO or 100 µM BNPP and incubated with 100 µM DPE for indicated time periods in serum-free MEM-α. The DPE-tagged cellular proteins were detected by HRP-streptavidin. (B) Structural comparison of DBE with DOPAC and DPE. (C) Effect of the pre-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in Hepa1c1c7 cells. Hepa1c1c7 cells were pre-incubated with DOPAC (open circle) or DBE (closed circle) for 30 min and incubated with 25 µM DPE for 3 h in serum-free MEM-α. (D) Effect of the co-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in cell lysate. Cell lysate was incubated with 25 µM DPE in combination with DOPAC (open circle) or DBE (closed circle) in the presence of 30 U laccase for 1 h. The values represent means ±S.D. of three separate experiments.

    Journal: Biochemistry and Biophysics Reports

    Article Title: A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite

    doi: 10.1016/j.bbrep.2016.06.020

    Figure Lengend Snippet: Detection of the intracellular DPE-modified proteins. (A) Detection of the DPE-modified proteins in Hepa1c1c7. Confluent Hepa1c1c7 cells were pre-incubated with 0.1% DMSO or 100 µM BNPP and incubated with 100 µM DPE for indicated time periods in serum-free MEM-α. The DPE-tagged cellular proteins were detected by HRP-streptavidin. (B) Structural comparison of DBE with DOPAC and DPE. (C) Effect of the pre-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in Hepa1c1c7 cells. Hepa1c1c7 cells were pre-incubated with DOPAC (open circle) or DBE (closed circle) for 30 min and incubated with 25 µM DPE for 3 h in serum-free MEM-α. (D) Effect of the co-treatment of DOPAC or DBE with DPE on the DPE-modified protein formation in cell lysate. Cell lysate was incubated with 25 µM DPE in combination with DOPAC (open circle) or DBE (closed circle) in the presence of 30 U laccase for 1 h. The values represent means ±S.D. of three separate experiments.

    Article Snippet: Streptavidin, HRP conjugate was purchased from Funakoshi (Tokyo, Japan).

    Techniques: Modification, Incubation

    CuAAC reaction of DPE with an azide-linked tag molecule. (A) Formation of 1,2,3-triazole by CuAAC reaction with DPE and benzyl azide. (B) Detection of GAPDH tagged by CuAAC reaction with DPE and the azide-labeled biotin. GAPDH (1 mg/ml) was incubated with or without 50 µM DPE in the presence or absence of 30 U laccase in 70 mM sodium phosphate buffer (pH 7.2) for 1 h at 37 °C. DPE-tagged GAPDH was detected by CBB staining (left) and HRP-streptavidin (right).

    Journal: Biochemistry and Biophysics Reports

    Article Title: A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite

    doi: 10.1016/j.bbrep.2016.06.020

    Figure Lengend Snippet: CuAAC reaction of DPE with an azide-linked tag molecule. (A) Formation of 1,2,3-triazole by CuAAC reaction with DPE and benzyl azide. (B) Detection of GAPDH tagged by CuAAC reaction with DPE and the azide-labeled biotin. GAPDH (1 mg/ml) was incubated with or without 50 µM DPE in the presence or absence of 30 U laccase in 70 mM sodium phosphate buffer (pH 7.2) for 1 h at 37 °C. DPE-tagged GAPDH was detected by CBB staining (left) and HRP-streptavidin (right).

    Article Snippet: Streptavidin, HRP conjugate was purchased from Funakoshi (Tokyo, Japan).

    Techniques: Labeling, Incubation, Staining

    Identification of the target proteins of DOPAC. Detection of the DPE-modified Keap1 and AhR. Confluent Hepa1c1c7 cells were incubated with 50 µM DPE for 5 h in serum-free MEM-α. The cell lysate was incubated with Streptavidin Mag Sepharose beads for 30 min. The DPE-modified Keap1 (left) or AhR (right) were detected by immunoblot analysis.

    Journal: Biochemistry and Biophysics Reports

    Article Title: A novel tag-free probe for targeting molecules interacting with a flavonoid catabolite

    doi: 10.1016/j.bbrep.2016.06.020

    Figure Lengend Snippet: Identification of the target proteins of DOPAC. Detection of the DPE-modified Keap1 and AhR. Confluent Hepa1c1c7 cells were incubated with 50 µM DPE for 5 h in serum-free MEM-α. The cell lysate was incubated with Streptavidin Mag Sepharose beads for 30 min. The DPE-modified Keap1 (left) or AhR (right) were detected by immunoblot analysis.

    Article Snippet: Streptavidin, HRP conjugate was purchased from Funakoshi (Tokyo, Japan).

    Techniques: Modification, Incubation

    Reactivity of mAbs against apoA-I in the lipid-free state or on reconstituted discoidal HDL by ELISA. Biotinylated Cys-apoA-I in the lipid-free state or on discoidal HDL were added to 96-well microplates coated with streptavidin. After washing, Ab#19-17, Ab#20-7, Ab#124-2, or Ab#187-2 were added to the plates with increasing concentrations, and then detected with POD-conjugated anti-mouse IgG antibody.

    Journal: Scientific Reports

    Article Title: Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies

    doi: 10.1038/s41598-017-03208-8

    Figure Lengend Snippet: Reactivity of mAbs against apoA-I in the lipid-free state or on reconstituted discoidal HDL by ELISA. Biotinylated Cys-apoA-I in the lipid-free state or on discoidal HDL were added to 96-well microplates coated with streptavidin. After washing, Ab#19-17, Ab#20-7, Ab#124-2, or Ab#187-2 were added to the plates with increasing concentrations, and then detected with POD-conjugated anti-mouse IgG antibody.

    Article Snippet: Single-antibody ELISA Microwells of the Costar microplates were coated overnight at 4 °C with 25 μg/ml streptavidin (Funakoshi, Tokyo, Japan) in PBS (100 μl/well), and then blocked by incubating with 1% Block Ace (DS Pharma Biomedical, Osaka, Japan) for 60 min at 37 °C.

    Techniques: Enzyme-linked Immunosorbent Assay

    Sandwich ELISA for detection of lipid-free and HDL-bound apoA-I. ApoA-I in the lipid-free state ( A ), on reconstituted discoidal HDL ( B ) and spherical HDL ( C ), and human plasma HDL ( D ) were added to the plates coated with Ab#124-2. After washing, biotinylated Ab#19-17, Ab#20-7, or Ab#187-2 were added to the plates, and then detected with POD-conjugated streptavidin.

    Journal: Scientific Reports

    Article Title: Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies

    doi: 10.1038/s41598-017-03208-8

    Figure Lengend Snippet: Sandwich ELISA for detection of lipid-free and HDL-bound apoA-I. ApoA-I in the lipid-free state ( A ), on reconstituted discoidal HDL ( B ) and spherical HDL ( C ), and human plasma HDL ( D ) were added to the plates coated with Ab#124-2. After washing, biotinylated Ab#19-17, Ab#20-7, or Ab#187-2 were added to the plates, and then detected with POD-conjugated streptavidin.

    Article Snippet: Single-antibody ELISA Microwells of the Costar microplates were coated overnight at 4 °C with 25 μg/ml streptavidin (Funakoshi, Tokyo, Japan) in PBS (100 μl/well), and then blocked by incubating with 1% Block Ace (DS Pharma Biomedical, Osaka, Japan) for 60 min at 37 °C.

    Techniques: Sandwich ELISA

    Reactivity of the present monoclonal antibodies to full-length and the N-terminal 1–83 fragment of apoA-I. Biotinylated full-length apoA-I or its N-terminal 1–83 fragment (0.3 μM) in G-PBS, or PBS (for examining the background value) was added to 96-well microplates coated with streptavidin, and then incubated for 30 min at 37 °C. After washing the wells with T-PBS, hybridoma supernatants, 500-fold diluted with G-PBS, were added to the wells (100 μl/well) and incubated at 37 °C for 30 min. After washing, POD-conjugated anti-mouse IgG antibody diluted at 1:5000 in G-PBS (100 μl/well) was added and incubated for 30 min at 37 °C. The wells were washed and bound POD activity was determined colorimetrically (see Materials and Methods). *** P

    Journal: Scientific Reports

    Article Title: Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies

    doi: 10.1038/s41598-017-03208-8

    Figure Lengend Snippet: Reactivity of the present monoclonal antibodies to full-length and the N-terminal 1–83 fragment of apoA-I. Biotinylated full-length apoA-I or its N-terminal 1–83 fragment (0.3 μM) in G-PBS, or PBS (for examining the background value) was added to 96-well microplates coated with streptavidin, and then incubated for 30 min at 37 °C. After washing the wells with T-PBS, hybridoma supernatants, 500-fold diluted with G-PBS, were added to the wells (100 μl/well) and incubated at 37 °C for 30 min. After washing, POD-conjugated anti-mouse IgG antibody diluted at 1:5000 in G-PBS (100 μl/well) was added and incubated for 30 min at 37 °C. The wells were washed and bound POD activity was determined colorimetrically (see Materials and Methods). *** P

    Article Snippet: Single-antibody ELISA Microwells of the Costar microplates were coated overnight at 4 °C with 25 μg/ml streptavidin (Funakoshi, Tokyo, Japan) in PBS (100 μl/well), and then blocked by incubating with 1% Block Ace (DS Pharma Biomedical, Osaka, Japan) for 60 min at 37 °C.

    Techniques: Incubation, Activity Assay