goat anti mouse horseradish peroxidase hrp light chain secondary antibody  (Abcam)

 
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    Name:
    Goat F ab 2 Anti Mouse IgM mu chain HRP
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    AB5930
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    Structured Review

    Abcam goat anti mouse horseradish peroxidase hrp light chain secondary antibody
    Detection of gK and UL20 on gradient-purified virions. Western <t>immunoblot</t> analysis of double-gradient-purified virions was performed using anti-V5 (gK) and anti-FLAG (UL20) antibodies. Individual lanes are labeled as follows: C, positive control for GAPDH; W, lysates from purified wild-type virus [HSV-1(F)-YE102]; M, lysates from purified mutant virus YE102-VC1; L, lysates from infected Vero cells; V, samples derived from purified virions. Individual blots were probed with anti-GAPDH, anti-ICP8, anti-gB, anti-gD, anti-gC, anti-FLAG (UL20), or anti-V5 (gK) antibody as indicated. The panel labeled at the top as IP:V5Ab shows immunoblots of immunoprecipitations with anti-V5 (gK) antibody probed with either anti-FLAG (UL20) or anti-V5 (gK) antibodies. In this panel, lanes are immunoprecipitates from mock-infected Vero cells (U), HSV-1(F)-YE102 purified virions (W), or YE102-VC1 purified virions (M). Molecular mass standards are shown with dots on each panel (250, 150, 100, 75, 50, 37, 25, 20, 15, and 10 kDa; Precision Plus protein standards; Bio-Rad). <t>HRP-conjugated</t> goat anti-mouse (HRP-GAb; IgG) was used for all data, except data shown on the panel labeled IP:V5Ab, wherein the F(ab) 2 - and Fc-purified portions of the HRP-GAb IgG were used for the α-V5 and α-FLAG panels, respectively.

    https://www.bioz.com/result/goat anti mouse horseradish peroxidase hrp light chain secondary antibody/product/Abcam
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse horseradish peroxidase hrp light chain secondary antibody - by Bioz Stars, 2021-06
    90/100 stars

    Images

    1) Product Images from "Site-Specific Proteolytic Cleavage of the Amino Terminus of Herpes Simplex Virus Glycoprotein K on Virion Particles Inhibits Virus Entry ▿"

    Article Title: Site-Specific Proteolytic Cleavage of the Amino Terminus of Herpes Simplex Virus Glycoprotein K on Virion Particles Inhibits Virus Entry ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.06268-11

    Detection of gK and UL20 on gradient-purified virions. Western immunoblot analysis of double-gradient-purified virions was performed using anti-V5 (gK) and anti-FLAG (UL20) antibodies. Individual lanes are labeled as follows: C, positive control for GAPDH; W, lysates from purified wild-type virus [HSV-1(F)-YE102]; M, lysates from purified mutant virus YE102-VC1; L, lysates from infected Vero cells; V, samples derived from purified virions. Individual blots were probed with anti-GAPDH, anti-ICP8, anti-gB, anti-gD, anti-gC, anti-FLAG (UL20), or anti-V5 (gK) antibody as indicated. The panel labeled at the top as IP:V5Ab shows immunoblots of immunoprecipitations with anti-V5 (gK) antibody probed with either anti-FLAG (UL20) or anti-V5 (gK) antibodies. In this panel, lanes are immunoprecipitates from mock-infected Vero cells (U), HSV-1(F)-YE102 purified virions (W), or YE102-VC1 purified virions (M). Molecular mass standards are shown with dots on each panel (250, 150, 100, 75, 50, 37, 25, 20, 15, and 10 kDa; Precision Plus protein standards; Bio-Rad). HRP-conjugated goat anti-mouse (HRP-GAb; IgG) was used for all data, except data shown on the panel labeled IP:V5Ab, wherein the F(ab) 2 - and Fc-purified portions of the HRP-GAb IgG were used for the α-V5 and α-FLAG panels, respectively.
    Figure Legend Snippet: Detection of gK and UL20 on gradient-purified virions. Western immunoblot analysis of double-gradient-purified virions was performed using anti-V5 (gK) and anti-FLAG (UL20) antibodies. Individual lanes are labeled as follows: C, positive control for GAPDH; W, lysates from purified wild-type virus [HSV-1(F)-YE102]; M, lysates from purified mutant virus YE102-VC1; L, lysates from infected Vero cells; V, samples derived from purified virions. Individual blots were probed with anti-GAPDH, anti-ICP8, anti-gB, anti-gD, anti-gC, anti-FLAG (UL20), or anti-V5 (gK) antibody as indicated. The panel labeled at the top as IP:V5Ab shows immunoblots of immunoprecipitations with anti-V5 (gK) antibody probed with either anti-FLAG (UL20) or anti-V5 (gK) antibodies. In this panel, lanes are immunoprecipitates from mock-infected Vero cells (U), HSV-1(F)-YE102 purified virions (W), or YE102-VC1 purified virions (M). Molecular mass standards are shown with dots on each panel (250, 150, 100, 75, 50, 37, 25, 20, 15, and 10 kDa; Precision Plus protein standards; Bio-Rad). HRP-conjugated goat anti-mouse (HRP-GAb; IgG) was used for all data, except data shown on the panel labeled IP:V5Ab, wherein the F(ab) 2 - and Fc-purified portions of the HRP-GAb IgG were used for the α-V5 and α-FLAG panels, respectively.

    Techniques Used: Purification, Western Blot, Labeling, Positive Control, Mutagenesis, Infection, Derivative Assay

    Related Articles

    Staining:

    Article Title: Laminin receptor initiates bacterial contact with the blood brain barrier in experimental meningitis models
    Article Snippet: For immunohistochemical staining for LR with and without infection, mice were challenged with PBS or 107 T4 pneumococci intranasally, and pneumonia was allowed to develop over 48 hours. .. Lungs were harvested, and frozen sections were stained with mouse anti-LR antibody MLuC5 (ab3099; Abcam) for 1 hour, followed by detection of binding with HRP-conjugated goat anti-mouse IgM (ab5930; Abcam) and the VIP Substrate Kit (Vector Laboratories). .. Bacterial LR-binding proteins were purified as previously described for the H. pylori SabA adhesion protein , with some modifications.

    Binding Assay:

    Article Title: Laminin receptor initiates bacterial contact with the blood brain barrier in experimental meningitis models
    Article Snippet: For immunohistochemical staining for LR with and without infection, mice were challenged with PBS or 107 T4 pneumococci intranasally, and pneumonia was allowed to develop over 48 hours. .. Lungs were harvested, and frozen sections were stained with mouse anti-LR antibody MLuC5 (ab3099; Abcam) for 1 hour, followed by detection of binding with HRP-conjugated goat anti-mouse IgM (ab5930; Abcam) and the VIP Substrate Kit (Vector Laboratories). .. Bacterial LR-binding proteins were purified as previously described for the H. pylori SabA adhesion protein , with some modifications.

    Article Title: Systemic autoimmunity induced by the TLR7/8 agonist Resiquimod causes myocarditis and dilated cardiomyopathy in a new mouse model of autoimmune heart disease
    Article Snippet: Serum from Resiquimod-treated mice was used in serial dilutions (1:10, 1:100, 1:1000) in triplicate for relative quantification of antibodies against myosin and troponin. .. Secondary HRP-conjugated goat anti-mouse IgM (ab5930), IgG1 (ab97240), IgG2a (ab97245) or IgG2b (ab97250) all from Abcam were used at 1:10,000, and TMB Substrate (BioLegend) was used to detect binding. .. For relative quantification of antibody isotypes in the serum, a Pierce Rapid Antibody Isotyping Kit - Mouse (Thermo Fisher Scientific) was used according to the manufacturer's instructions, using previously determined dilutions optimised for individual isotypes ( Fig. S3 ).

    Incubation:

    Article Title: Functional Compartmentalization of the Plasma Membrane of Neurons by a Unique Acyl Chain Composition of Phospholipids *
    Article Snippet: For binding measurements to lipid standards, the plates were blocked with 0.1% ovalbumin (Wako) in PBS for 1 h and then incubated for 30 min with 10 μg/liter mAb#15 antibody in 5% milk, 3% normal goat serum, PBS. .. After washing with PBS, the secondary antibody (HRP-conjugated F(ab′)2 antibody fragment to mouse IgM (ab5930, Abcam), 2,000-fold dilution in 1% BSA, PBS) was added, followed by incubation for 30 min. After washing the plates with PBS, immunoreactivity was quantified as above. .. Dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine (DPPC) and sphingomyelin from bovine brain (S7004) were from Sigma.

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  • 99
    Abcam goat anti mouse hrp secondary antibody
    Conjugation efficiency and biodistribution analysis of CRISPR-modified antibody. ( a ) Coomassie blue stained gel of sortase-mediated conjugation of modified antibody to GGG-fluorophore. ( b ) Fluorescence scan of the antibody-fluorophore, stained with <t>anti-Flag-HRP,</t> showing the antibody conjugated fluorophore (red tab) compared to free fluorophore (black tab). ( c ) HPLC fluorescence trace antibody-fluorophore conjugates at different molar ratios. ( d ) Quantitative analysis of the percent of antibody modification. ( e ) Binding of 125 I-labeled CRISPR-modified antibody to REN-ICAM and REN-WT cells. ( f ) Biodistribution of 125 I-labeled CRISPR-modified anti-ICAM mAb in mice at 30 min. Tissue uptake is indicated as mean ± SEM (n = 3). Biodistribution analysis was performed comparing 111 In-labeled site-specific CRISPR/Cas9- to chemically-modified mAb. ( g ) Biodistribution of 111 In-labeled anti-ICAM mAb modified in mice at 30 min. Tissue uptake is indicated as mean ± SEM. ( h ) Localization ratio of selected organs. Significant differences were determined by t-test with Bonferroni correction to account for multiple comparisons.
    Goat Anti Mouse Hrp Secondary Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse hrp secondary antibody/product/Abcam
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse hrp secondary antibody - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

    90
    Abcam goat anti mouse horseradish peroxidase hrp light chain secondary antibody
    Detection of gK and UL20 on gradient-purified virions. Western <t>immunoblot</t> analysis of double-gradient-purified virions was performed using anti-V5 (gK) and anti-FLAG (UL20) antibodies. Individual lanes are labeled as follows: C, positive control for GAPDH; W, lysates from purified wild-type virus [HSV-1(F)-YE102]; M, lysates from purified mutant virus YE102-VC1; L, lysates from infected Vero cells; V, samples derived from purified virions. Individual blots were probed with anti-GAPDH, anti-ICP8, anti-gB, anti-gD, anti-gC, anti-FLAG (UL20), or anti-V5 (gK) antibody as indicated. The panel labeled at the top as IP:V5Ab shows immunoblots of immunoprecipitations with anti-V5 (gK) antibody probed with either anti-FLAG (UL20) or anti-V5 (gK) antibodies. In this panel, lanes are immunoprecipitates from mock-infected Vero cells (U), HSV-1(F)-YE102 purified virions (W), or YE102-VC1 purified virions (M). Molecular mass standards are shown with dots on each panel (250, 150, 100, 75, 50, 37, 25, 20, 15, and 10 kDa; Precision Plus protein standards; Bio-Rad). <t>HRP-conjugated</t> goat anti-mouse (HRP-GAb; IgG) was used for all data, except data shown on the panel labeled IP:V5Ab, wherein the F(ab) 2 - and Fc-purified portions of the HRP-GAb IgG were used for the α-V5 and α-FLAG panels, respectively.
    Goat Anti Mouse Horseradish Peroxidase Hrp Light Chain Secondary Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse horseradish peroxidase hrp light chain secondary antibody/product/Abcam
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse horseradish peroxidase hrp light chain secondary antibody - by Bioz Stars, 2021-06
    90/100 stars
      Buy from Supplier

    96
    Abcam goat anti mouse igg h l hrp secondary antibodies
    Detection of gK and UL20 on gradient-purified virions. Western <t>immunoblot</t> analysis of double-gradient-purified virions was performed using anti-V5 (gK) and anti-FLAG (UL20) antibodies. Individual lanes are labeled as follows: C, positive control for GAPDH; W, lysates from purified wild-type virus [HSV-1(F)-YE102]; M, lysates from purified mutant virus YE102-VC1; L, lysates from infected Vero cells; V, samples derived from purified virions. Individual blots were probed with anti-GAPDH, anti-ICP8, anti-gB, anti-gD, anti-gC, anti-FLAG (UL20), or anti-V5 (gK) antibody as indicated. The panel labeled at the top as IP:V5Ab shows immunoblots of immunoprecipitations with anti-V5 (gK) antibody probed with either anti-FLAG (UL20) or anti-V5 (gK) antibodies. In this panel, lanes are immunoprecipitates from mock-infected Vero cells (U), HSV-1(F)-YE102 purified virions (W), or YE102-VC1 purified virions (M). Molecular mass standards are shown with dots on each panel (250, 150, 100, 75, 50, 37, 25, 20, 15, and 10 kDa; Precision Plus protein standards; Bio-Rad). <t>HRP-conjugated</t> goat anti-mouse (HRP-GAb; IgG) was used for all data, except data shown on the panel labeled IP:V5Ab, wherein the F(ab) 2 - and Fc-purified portions of the HRP-GAb IgG were used for the α-V5 and α-FLAG panels, respectively.
    Goat Anti Mouse Igg H L Hrp Secondary Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse igg h l hrp secondary antibodies/product/Abcam
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg h l hrp secondary antibodies - by Bioz Stars, 2021-06
    96/100 stars
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    Image Search Results


    Conjugation efficiency and biodistribution analysis of CRISPR-modified antibody. ( a ) Coomassie blue stained gel of sortase-mediated conjugation of modified antibody to GGG-fluorophore. ( b ) Fluorescence scan of the antibody-fluorophore, stained with anti-Flag-HRP, showing the antibody conjugated fluorophore (red tab) compared to free fluorophore (black tab). ( c ) HPLC fluorescence trace antibody-fluorophore conjugates at different molar ratios. ( d ) Quantitative analysis of the percent of antibody modification. ( e ) Binding of 125 I-labeled CRISPR-modified antibody to REN-ICAM and REN-WT cells. ( f ) Biodistribution of 125 I-labeled CRISPR-modified anti-ICAM mAb in mice at 30 min. Tissue uptake is indicated as mean ± SEM (n = 3). Biodistribution analysis was performed comparing 111 In-labeled site-specific CRISPR/Cas9- to chemically-modified mAb. ( g ) Biodistribution of 111 In-labeled anti-ICAM mAb modified in mice at 30 min. Tissue uptake is indicated as mean ± SEM. ( h ) Localization ratio of selected organs. Significant differences were determined by t-test with Bonferroni correction to account for multiple comparisons.

    Journal: Scientific Reports

    Article Title: Molecular engineering of antibodies for site-specific covalent conjugation using CRISPR/Cas9

    doi: 10.1038/s41598-018-19784-2

    Figure Lengend Snippet: Conjugation efficiency and biodistribution analysis of CRISPR-modified antibody. ( a ) Coomassie blue stained gel of sortase-mediated conjugation of modified antibody to GGG-fluorophore. ( b ) Fluorescence scan of the antibody-fluorophore, stained with anti-Flag-HRP, showing the antibody conjugated fluorophore (red tab) compared to free fluorophore (black tab). ( c ) HPLC fluorescence trace antibody-fluorophore conjugates at different molar ratios. ( d ) Quantitative analysis of the percent of antibody modification. ( e ) Binding of 125 I-labeled CRISPR-modified antibody to REN-ICAM and REN-WT cells. ( f ) Biodistribution of 125 I-labeled CRISPR-modified anti-ICAM mAb in mice at 30 min. Tissue uptake is indicated as mean ± SEM (n = 3). Biodistribution analysis was performed comparing 111 In-labeled site-specific CRISPR/Cas9- to chemically-modified mAb. ( g ) Biodistribution of 111 In-labeled anti-ICAM mAb modified in mice at 30 min. Tissue uptake is indicated as mean ± SEM. ( h ) Localization ratio of selected organs. Significant differences were determined by t-test with Bonferroni correction to account for multiple comparisons.

    Article Snippet: The membrane was incubated with anti-FLAG M2 antibody (F3165; Sigma-Aldrich, St. Louis, MO), followed by goat anti-mouse HRP secondary antibody (ab6789; Abcam, Cambridge, MA).

    Techniques: Conjugation Assay, CRISPR, Modification, Staining, Fluorescence, High Performance Liquid Chromatography, Binding Assay, Labeling, Mouse Assay

    Schematic diagram and characterization of CRISPR/Cas9 genome editing of hybridoma cells for site-specific modification of antibodies. C-terminal end of Ig gamma-2B gene from ( a ) rat chromosome 13q24, ( b ) Fcgr2b locus, was modified by ( c ) homology directed repair to incorporate sortase and flag tags. sgRNAs with best On-Target/Off-Target scores were selected in the region near the IgG2b stop codon. ( d ) Anti-ICAMIgG2b antibodies were generated incorporating Sortase and FLAG tags at their C-terminal end. Confirmation of insert was performed by ( e ) Agarose gel analysis of PCR fragments from positive and negative clones. ( f ) Verification of integration in positive clones by sanger sequencing. ( g ) Coomassie blue stained gel of three positive clones. ( h ) Western blot of a positive clone under reducing and non-reducing conditions, stained with anti-Flag-HRP.

    Journal: Scientific Reports

    Article Title: Molecular engineering of antibodies for site-specific covalent conjugation using CRISPR/Cas9

    doi: 10.1038/s41598-018-19784-2

    Figure Lengend Snippet: Schematic diagram and characterization of CRISPR/Cas9 genome editing of hybridoma cells for site-specific modification of antibodies. C-terminal end of Ig gamma-2B gene from ( a ) rat chromosome 13q24, ( b ) Fcgr2b locus, was modified by ( c ) homology directed repair to incorporate sortase and flag tags. sgRNAs with best On-Target/Off-Target scores were selected in the region near the IgG2b stop codon. ( d ) Anti-ICAMIgG2b antibodies were generated incorporating Sortase and FLAG tags at their C-terminal end. Confirmation of insert was performed by ( e ) Agarose gel analysis of PCR fragments from positive and negative clones. ( f ) Verification of integration in positive clones by sanger sequencing. ( g ) Coomassie blue stained gel of three positive clones. ( h ) Western blot of a positive clone under reducing and non-reducing conditions, stained with anti-Flag-HRP.

    Article Snippet: The membrane was incubated with anti-FLAG M2 antibody (F3165; Sigma-Aldrich, St. Louis, MO), followed by goat anti-mouse HRP secondary antibody (ab6789; Abcam, Cambridge, MA).

    Techniques: CRISPR, Modification, Generated, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Clone Assay, Sequencing, Staining, Western Blot

    Conjugation efficiency and biodistribution analysis of CRISPR-modified antibody. ( a ) Coomassie blue stained gel of sortase-mediated conjugation of modified antibody to GGG-fluorophore. ( b ) Fluorescence scan of the antibody-fluorophore, stained with anti-Flag-HRP, showing the antibody conjugated fluorophore (red tab) compared to free fluorophore (black tab). ( c ) HPLC fluorescence trace antibody-fluorophore conjugates at different molar ratios. ( d ) Quantitative analysis of the percent of antibody modification. ( e ) Binding of 125 I-labeled CRISPR-modified antibody to REN-ICAM and REN-WT cells. ( f ) Biodistribution of 125 I-labeled CRISPR-modified anti-ICAM mAb in mice at 30 min. Tissue uptake is indicated as mean ± SEM (n = 3). Biodistribution analysis was performed comparing 111 In-labeled site-specific CRISPR/Cas9- to chemically-modified mAb. ( g ) Biodistribution of 111 In-labeled anti-ICAM mAb modified in mice at 30 min. Tissue uptake is indicated as mean ± SEM. ( h ) Localization ratio of selected organs. Significant differences were determined by t-test with Bonferroni correction to account for multiple comparisons.

    Journal: Scientific Reports

    Article Title: Molecular engineering of antibodies for site-specific covalent conjugation using CRISPR/Cas9

    doi: 10.1038/s41598-018-19784-2

    Figure Lengend Snippet: Conjugation efficiency and biodistribution analysis of CRISPR-modified antibody. ( a ) Coomassie blue stained gel of sortase-mediated conjugation of modified antibody to GGG-fluorophore. ( b ) Fluorescence scan of the antibody-fluorophore, stained with anti-Flag-HRP, showing the antibody conjugated fluorophore (red tab) compared to free fluorophore (black tab). ( c ) HPLC fluorescence trace antibody-fluorophore conjugates at different molar ratios. ( d ) Quantitative analysis of the percent of antibody modification. ( e ) Binding of 125 I-labeled CRISPR-modified antibody to REN-ICAM and REN-WT cells. ( f ) Biodistribution of 125 I-labeled CRISPR-modified anti-ICAM mAb in mice at 30 min. Tissue uptake is indicated as mean ± SEM (n = 3). Biodistribution analysis was performed comparing 111 In-labeled site-specific CRISPR/Cas9- to chemically-modified mAb. ( g ) Biodistribution of 111 In-labeled anti-ICAM mAb modified in mice at 30 min. Tissue uptake is indicated as mean ± SEM. ( h ) Localization ratio of selected organs. Significant differences were determined by t-test with Bonferroni correction to account for multiple comparisons.

    Article Snippet: The membrane was incubated with anti-FLAG M2 antibody (F3165; Sigma-Aldrich, St. Louis, MO), followed by goat anti-mouse HRP secondary antibody (ab6789; Abcam, Cambridge, MA).

    Techniques: Conjugation Assay, CRISPR, Modification, Staining, Fluorescence, High Performance Liquid Chromatography, Binding Assay, Labeling, Mouse Assay

    Schematic diagram and characterization of CRISPR/Cas9 genome editing of hybridoma cells for site-specific modification of antibodies. C-terminal end of Ig gamma-2B gene from ( a ) rat chromosome 13q24, ( b ) Fcgr2b locus, was modified by ( c ) homology directed repair to incorporate sortase and flag tags. sgRNAs with best On-Target/Off-Target scores were selected in the region near the IgG2b stop codon. ( d ) Anti-ICAMIgG2b antibodies were generated incorporating Sortase and FLAG tags at their C-terminal end. Confirmation of insert was performed by ( e ) Agarose gel analysis of PCR fragments from positive and negative clones. ( f ) Verification of integration in positive clones by sanger sequencing. ( g ) Coomassie blue stained gel of three positive clones. ( h ) Western blot of a positive clone under reducing and non-reducing conditions, stained with anti-Flag-HRP.

    Journal: Scientific Reports

    Article Title: Molecular engineering of antibodies for site-specific covalent conjugation using CRISPR/Cas9

    doi: 10.1038/s41598-018-19784-2

    Figure Lengend Snippet: Schematic diagram and characterization of CRISPR/Cas9 genome editing of hybridoma cells for site-specific modification of antibodies. C-terminal end of Ig gamma-2B gene from ( a ) rat chromosome 13q24, ( b ) Fcgr2b locus, was modified by ( c ) homology directed repair to incorporate sortase and flag tags. sgRNAs with best On-Target/Off-Target scores were selected in the region near the IgG2b stop codon. ( d ) Anti-ICAMIgG2b antibodies were generated incorporating Sortase and FLAG tags at their C-terminal end. Confirmation of insert was performed by ( e ) Agarose gel analysis of PCR fragments from positive and negative clones. ( f ) Verification of integration in positive clones by sanger sequencing. ( g ) Coomassie blue stained gel of three positive clones. ( h ) Western blot of a positive clone under reducing and non-reducing conditions, stained with anti-Flag-HRP.

    Article Snippet: The membrane was incubated with anti-FLAG M2 antibody (F3165; Sigma-Aldrich, St. Louis, MO), followed by goat anti-mouse HRP secondary antibody (ab6789; Abcam, Cambridge, MA).

    Techniques: CRISPR, Modification, Generated, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Clone Assay, Sequencing, Staining, Western Blot

    Detection of gK and UL20 on gradient-purified virions. Western immunoblot analysis of double-gradient-purified virions was performed using anti-V5 (gK) and anti-FLAG (UL20) antibodies. Individual lanes are labeled as follows: C, positive control for GAPDH; W, lysates from purified wild-type virus [HSV-1(F)-YE102]; M, lysates from purified mutant virus YE102-VC1; L, lysates from infected Vero cells; V, samples derived from purified virions. Individual blots were probed with anti-GAPDH, anti-ICP8, anti-gB, anti-gD, anti-gC, anti-FLAG (UL20), or anti-V5 (gK) antibody as indicated. The panel labeled at the top as IP:V5Ab shows immunoblots of immunoprecipitations with anti-V5 (gK) antibody probed with either anti-FLAG (UL20) or anti-V5 (gK) antibodies. In this panel, lanes are immunoprecipitates from mock-infected Vero cells (U), HSV-1(F)-YE102 purified virions (W), or YE102-VC1 purified virions (M). Molecular mass standards are shown with dots on each panel (250, 150, 100, 75, 50, 37, 25, 20, 15, and 10 kDa; Precision Plus protein standards; Bio-Rad). HRP-conjugated goat anti-mouse (HRP-GAb; IgG) was used for all data, except data shown on the panel labeled IP:V5Ab, wherein the F(ab) 2 - and Fc-purified portions of the HRP-GAb IgG were used for the α-V5 and α-FLAG panels, respectively.

    Journal: Journal of Virology

    Article Title: Site-Specific Proteolytic Cleavage of the Amino Terminus of Herpes Simplex Virus Glycoprotein K on Virion Particles Inhibits Virus Entry ▿

    doi: 10.1128/JVI.06268-11

    Figure Lengend Snippet: Detection of gK and UL20 on gradient-purified virions. Western immunoblot analysis of double-gradient-purified virions was performed using anti-V5 (gK) and anti-FLAG (UL20) antibodies. Individual lanes are labeled as follows: C, positive control for GAPDH; W, lysates from purified wild-type virus [HSV-1(F)-YE102]; M, lysates from purified mutant virus YE102-VC1; L, lysates from infected Vero cells; V, samples derived from purified virions. Individual blots were probed with anti-GAPDH, anti-ICP8, anti-gB, anti-gD, anti-gC, anti-FLAG (UL20), or anti-V5 (gK) antibody as indicated. The panel labeled at the top as IP:V5Ab shows immunoblots of immunoprecipitations with anti-V5 (gK) antibody probed with either anti-FLAG (UL20) or anti-V5 (gK) antibodies. In this panel, lanes are immunoprecipitates from mock-infected Vero cells (U), HSV-1(F)-YE102 purified virions (W), or YE102-VC1 purified virions (M). Molecular mass standards are shown with dots on each panel (250, 150, 100, 75, 50, 37, 25, 20, 15, and 10 kDa; Precision Plus protein standards; Bio-Rad). HRP-conjugated goat anti-mouse (HRP-GAb; IgG) was used for all data, except data shown on the panel labeled IP:V5Ab, wherein the F(ab) 2 - and Fc-purified portions of the HRP-GAb IgG were used for the α-V5 and α-FLAG panels, respectively.

    Article Snippet: Immunoblot assays were carried out using mouse anti-V5 antibody and goat anti-mouse horseradish peroxidase (HRP) light-chain secondary antibody (Abcam, Inc., Cambridge, MA).

    Techniques: Purification, Western Blot, Labeling, Positive Control, Mutagenesis, Infection, Derivative Assay