peroxidase conjugated secondary antibody  (Thermo Fisher)


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    Thermo Fisher peroxidase conjugated secondary antibody
    Interaction of SARS-CoV-2 S protein with N protein and effects of R-Spike CD-SARS-CoV-2 on production of SARS-CoV-2 proteins. (A) Interaction of SARS-CoV-2 S protein with N protein. Lysates were prepared from uninfected and SARS-CoV-2 (0.1 MOI)-infected Vero cells. The lysates were immunoprecipitated with anti-SARS-CoV-2 S mAb (left). The immunocomplexes were subjected to western blotting with anti-SARS-CoV-2 S mAb or anti-SARS-CoV-2 N Ab. The cell lysates were analyzed by western blotting with the indicated antibodies (right). Anti-S Ab, anti-SARS-CoV-2 S Ab. Anti-N mAb, anti-SARS-CoV-2 N mAb. (B-E) Effects of R-Spike CD-SARS-CoV-2 on production of SARS-CoV-2 proteins. Vero cells (B and C) and Calu-3 cells (D and E) were infected with SARS-CoV-2 (0.1 MOI) and then treated with PBS or 2 μM of cell-penetrating peptides (R-Spike CD-SARS-CoV-2 or R-CP-1) at 6 h after virus infection (n = 3) in DMEM medium containing 2% FBS. The cells were cultured for 48 h and then analyzed by confocal microscopy after staining with anti-SARS-CoV-2 S Ab (B and D) or anti-SARS-CoV-2 N mAb (C and E) and then, Alexa Fluor <t>488-conjugated</t> <t>secondary</t> <t>antibody.</t> Scale bar, 20 μm. These results are representative of two independent experiments.
    Peroxidase Conjugated Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99/100 stars

    Images

    1) Product Images from "MERS-CoV and SARS-CoV-2 replication can be inhibited by targeting the interaction between the viral spike protein and the nucleocapsid protein"

    Article Title: MERS-CoV and SARS-CoV-2 replication can be inhibited by targeting the interaction between the viral spike protein and the nucleocapsid protein

    Journal: Theranostics

    doi: 10.7150/thno.55647

    Interaction of SARS-CoV-2 S protein with N protein and effects of R-Spike CD-SARS-CoV-2 on production of SARS-CoV-2 proteins. (A) Interaction of SARS-CoV-2 S protein with N protein. Lysates were prepared from uninfected and SARS-CoV-2 (0.1 MOI)-infected Vero cells. The lysates were immunoprecipitated with anti-SARS-CoV-2 S mAb (left). The immunocomplexes were subjected to western blotting with anti-SARS-CoV-2 S mAb or anti-SARS-CoV-2 N Ab. The cell lysates were analyzed by western blotting with the indicated antibodies (right). Anti-S Ab, anti-SARS-CoV-2 S Ab. Anti-N mAb, anti-SARS-CoV-2 N mAb. (B-E) Effects of R-Spike CD-SARS-CoV-2 on production of SARS-CoV-2 proteins. Vero cells (B and C) and Calu-3 cells (D and E) were infected with SARS-CoV-2 (0.1 MOI) and then treated with PBS or 2 μM of cell-penetrating peptides (R-Spike CD-SARS-CoV-2 or R-CP-1) at 6 h after virus infection (n = 3) in DMEM medium containing 2% FBS. The cells were cultured for 48 h and then analyzed by confocal microscopy after staining with anti-SARS-CoV-2 S Ab (B and D) or anti-SARS-CoV-2 N mAb (C and E) and then, Alexa Fluor 488-conjugated secondary antibody. Scale bar, 20 μm. These results are representative of two independent experiments.
    Figure Legend Snippet: Interaction of SARS-CoV-2 S protein with N protein and effects of R-Spike CD-SARS-CoV-2 on production of SARS-CoV-2 proteins. (A) Interaction of SARS-CoV-2 S protein with N protein. Lysates were prepared from uninfected and SARS-CoV-2 (0.1 MOI)-infected Vero cells. The lysates were immunoprecipitated with anti-SARS-CoV-2 S mAb (left). The immunocomplexes were subjected to western blotting with anti-SARS-CoV-2 S mAb or anti-SARS-CoV-2 N Ab. The cell lysates were analyzed by western blotting with the indicated antibodies (right). Anti-S Ab, anti-SARS-CoV-2 S Ab. Anti-N mAb, anti-SARS-CoV-2 N mAb. (B-E) Effects of R-Spike CD-SARS-CoV-2 on production of SARS-CoV-2 proteins. Vero cells (B and C) and Calu-3 cells (D and E) were infected with SARS-CoV-2 (0.1 MOI) and then treated with PBS or 2 μM of cell-penetrating peptides (R-Spike CD-SARS-CoV-2 or R-CP-1) at 6 h after virus infection (n = 3) in DMEM medium containing 2% FBS. The cells were cultured for 48 h and then analyzed by confocal microscopy after staining with anti-SARS-CoV-2 S Ab (B and D) or anti-SARS-CoV-2 N mAb (C and E) and then, Alexa Fluor 488-conjugated secondary antibody. Scale bar, 20 μm. These results are representative of two independent experiments.

    Techniques Used: Infection, Immunoprecipitation, Western Blot, Cell Culture, Confocal Microscopy, Staining

    2) Product Images from "Herpes Simplex Virus Glycoprotein C Regulates Low pH Entry"

    Article Title: Herpes Simplex Virus Glycoprotein C Regulates Low pH Entry

    Journal: bioRxiv

    doi: 10.1101/858472

    HSV-1 gC facilitates acid-induced conformational changes in the fusion protein gB. Extracellular preparations of HSV-1 gCR or ΔgC (∼ 10 7 genome copy numbers) were treated with pHs ranging from 7.3 to 5.0 and blotted directly to nitrocellulose. Blots were probed with the indicated gB MAbs H126, H1838, SS144, or H1359 at neutral pH followed by HRP-conjugated anti-mouse secondary antibody. The antibody name is shown at the left and the gB domain to which each MAb is directed is indicated in parenthesis. These are individual examples of experiments that were quantitated and averaged together with multiple similar independent determinations. Summarized quantitative results are depicted in Figure 6 .
    Figure Legend Snippet: HSV-1 gC facilitates acid-induced conformational changes in the fusion protein gB. Extracellular preparations of HSV-1 gCR or ΔgC (∼ 10 7 genome copy numbers) were treated with pHs ranging from 7.3 to 5.0 and blotted directly to nitrocellulose. Blots were probed with the indicated gB MAbs H126, H1838, SS144, or H1359 at neutral pH followed by HRP-conjugated anti-mouse secondary antibody. The antibody name is shown at the left and the gB domain to which each MAb is directed is indicated in parenthesis. These are individual examples of experiments that were quantitated and averaged together with multiple similar independent determinations. Summarized quantitative results are depicted in Figure 6 .

    Techniques Used:

    3) Product Images from "Docking of acetyl-CoA carboxylase to the plastid envelope membrane attenuates fatty acid production in plants"

    Article Title: Docking of acetyl-CoA carboxylase to the plastid envelope membrane attenuates fatty acid production in plants

    Journal: Nature Communications

    doi: 10.1038/s41467-020-20014-5

    CTIs are localized in the inner envelope membrane of chloroplasts. a Localization of CTIs and α-CT in Arabidopsis protoplasts. CTIs and α-CT were fused with YFP, and expressed in protoplasts. The fluorescence was detected by confocal microscopy 16 h after transformation. Empty vector was used as a control. Experiments were repeated three times with similar results. For each of the three independent protoplast transformations, at least five different protoplasts were observed. Bars = 5 µm. b Colocalization of CTI1-GFP and α-CT-RFP fusions transiently coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. c Colocalization of CTI1-GFP and TGD2-RFP fusions coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. d Co-immunolocalization of CTI1 and the E37 marker of the inner envelope membrane of chloroplasts. Co-immunolocalization experiments were carried out in using anti-E37 primary antibodies and secondary antibodies conjugated to Alexa Fluor 568 (fluorescence observed at 603-682 nm; red signal) or specific anti-HA antibodies and secondary antibodies conjugated to Alexa Fluor 488 (fluorescence observed at 495–585 nm; green signal). Merged pictures are also presented. Experiments with protoplasts were repeated three times with similar results. For each of the three independent protoplast transformations, eight different protoplasts were observed. Experiments with embryos were repeated twice with similar results. In each experiment, 10 different cells from distinct embryos were observed. Bars = 5 µm. e Topology of the CTI1 protein. Intact Arabidopsis chloroplasts were digested by thermolysin (Thr) or trypsin (Typ) for 30 min. Proteins were detected with anti-CTI1 or anti-α-CT antibodies. TOC33-MYC fusion protein, used as a control, was detected with anti-MYC antibodies. The results shown are representative of three biologically independent samples.
    Figure Legend Snippet: CTIs are localized in the inner envelope membrane of chloroplasts. a Localization of CTIs and α-CT in Arabidopsis protoplasts. CTIs and α-CT were fused with YFP, and expressed in protoplasts. The fluorescence was detected by confocal microscopy 16 h after transformation. Empty vector was used as a control. Experiments were repeated three times with similar results. For each of the three independent protoplast transformations, at least five different protoplasts were observed. Bars = 5 µm. b Colocalization of CTI1-GFP and α-CT-RFP fusions transiently coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. c Colocalization of CTI1-GFP and TGD2-RFP fusions coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. d Co-immunolocalization of CTI1 and the E37 marker of the inner envelope membrane of chloroplasts. Co-immunolocalization experiments were carried out in using anti-E37 primary antibodies and secondary antibodies conjugated to Alexa Fluor 568 (fluorescence observed at 603-682 nm; red signal) or specific anti-HA antibodies and secondary antibodies conjugated to Alexa Fluor 488 (fluorescence observed at 495–585 nm; green signal). Merged pictures are also presented. Experiments with protoplasts were repeated three times with similar results. For each of the three independent protoplast transformations, eight different protoplasts were observed. Experiments with embryos were repeated twice with similar results. In each experiment, 10 different cells from distinct embryos were observed. Bars = 5 µm. e Topology of the CTI1 protein. Intact Arabidopsis chloroplasts were digested by thermolysin (Thr) or trypsin (Typ) for 30 min. Proteins were detected with anti-CTI1 or anti-α-CT antibodies. TOC33-MYC fusion protein, used as a control, was detected with anti-MYC antibodies. The results shown are representative of three biologically independent samples.

    Techniques Used: Fluorescence, Confocal Microscopy, Transformation Assay, Plasmid Preparation, Marker

    4) Product Images from "Docking of acetyl-CoA carboxylase to the plastid envelope membrane attenuates fatty acid production in plants"

    Article Title: Docking of acetyl-CoA carboxylase to the plastid envelope membrane attenuates fatty acid production in plants

    Journal: Nature Communications

    doi: 10.1038/s41467-020-20014-5

    CTIs are localized in the inner envelope membrane of chloroplasts. a Localization of CTIs and α-CT in Arabidopsis protoplasts. CTIs and α-CT were fused with YFP, and expressed in protoplasts. The fluorescence was detected by confocal microscopy 16 h after transformation. Empty vector was used as a control. Experiments were repeated three times with similar results. For each of the three independent protoplast transformations, at least five different protoplasts were observed. Bars = 5 µm. b Colocalization of CTI1-GFP and α-CT-RFP fusions transiently coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. c Colocalization of CTI1-GFP and TGD2-RFP fusions coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. d Co-immunolocalization of CTI1 and the E37 marker of the inner envelope membrane of chloroplasts. Co-immunolocalization experiments were carried out in using anti-E37 primary antibodies and secondary antibodies conjugated to Alexa Fluor 568 (fluorescence observed at 603-682 nm; red signal) or specific anti-HA antibodies and secondary antibodies conjugated to Alexa Fluor 488 (fluorescence observed at 495–585 nm; green signal). Merged pictures are also presented. Experiments with protoplasts were repeated three times with similar results. For each of the three independent protoplast transformations, eight different protoplasts were observed. Experiments with embryos were repeated twice with similar results. In each experiment, 10 different cells from distinct embryos were observed. Bars = 5 µm. e Topology of the CTI1 protein. Intact Arabidopsis chloroplasts were digested by thermolysin (Thr) or trypsin (Typ) for 30 min. Proteins were detected with anti-CTI1 or anti-α-CT antibodies. TOC33-MYC fusion protein, used as a control, was detected with anti-MYC antibodies. The results shown are representative of three biologically independent samples.
    Figure Legend Snippet: CTIs are localized in the inner envelope membrane of chloroplasts. a Localization of CTIs and α-CT in Arabidopsis protoplasts. CTIs and α-CT were fused with YFP, and expressed in protoplasts. The fluorescence was detected by confocal microscopy 16 h after transformation. Empty vector was used as a control. Experiments were repeated three times with similar results. For each of the three independent protoplast transformations, at least five different protoplasts were observed. Bars = 5 µm. b Colocalization of CTI1-GFP and α-CT-RFP fusions transiently coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. c Colocalization of CTI1-GFP and TGD2-RFP fusions coexpressed in leaves of Nicotiana benthamiana and observed two days after infiltration. Experiments were repeated three times with similar results. For each of the three independent transformations, at least five different cells were observed. Bars = 5 µm. d Co-immunolocalization of CTI1 and the E37 marker of the inner envelope membrane of chloroplasts. Co-immunolocalization experiments were carried out in using anti-E37 primary antibodies and secondary antibodies conjugated to Alexa Fluor 568 (fluorescence observed at 603-682 nm; red signal) or specific anti-HA antibodies and secondary antibodies conjugated to Alexa Fluor 488 (fluorescence observed at 495–585 nm; green signal). Merged pictures are also presented. Experiments with protoplasts were repeated three times with similar results. For each of the three independent protoplast transformations, eight different protoplasts were observed. Experiments with embryos were repeated twice with similar results. In each experiment, 10 different cells from distinct embryos were observed. Bars = 5 µm. e Topology of the CTI1 protein. Intact Arabidopsis chloroplasts were digested by thermolysin (Thr) or trypsin (Typ) for 30 min. Proteins were detected with anti-CTI1 or anti-α-CT antibodies. TOC33-MYC fusion protein, used as a control, was detected with anti-MYC antibodies. The results shown are representative of three biologically independent samples.

    Techniques Used: Fluorescence, Confocal Microscopy, Transformation Assay, Plasmid Preparation, Marker

    5) Product Images from "Zika virus antagonizes interferon response in patients and disrupts RIG-I–MAVS interaction through its CARD-TM domains"

    Article Title: Zika virus antagonizes interferon response in patients and disrupts RIG-I–MAVS interaction through its CARD-TM domains

    Journal: Cell & Bioscience

    doi: 10.1186/s13578-019-0308-9

    ZIKV NS4A co-localizes and interacts with MAVS. a HeLa cells transfected with plasmids expressing Flag-tagged NS4A, Flag-tagged ZIKV prM or influenza Flag-tagged PB1-F2 were stained with anti-Flag and anti-MAVS antibodies as well as DAPI. Secondary antibodies conjugated to rhodamine and FITC dye were used to visualize the indicated proteins. Images are representative of three independent experiments. 293T cells co-transfected with plasmids encoding Myc-tagged MAVS and Flag-tagged NS4A were used in a co-IP assay to address whether ZIKV NS4A protein physically interacts with MAVS. Cell lysates were precipitated with an anti-Flag antibody ( b ), anti-Myc antibody ( c ), or control mouse IgG, and immunocomplexes were analyzed with the indicated antibodies by western blotting. d 293T cells were transfected with plasmids encoding Flag-tagged NS4A, followed by immunoprecipitation using anti-Flag antibody or control IgG. The immunocomplexes were analyzed with anti-MAVS antibody by Western blotting. e HFF-1 cells were infected with ZIKV at an MOI of 5 followed by immunoprecipitation using anti-MAVS antibody or control mouse IgG. The immunocomplexes that were captured by the protein G Dynabeads were analyzed by Western blotting using anti-NS4A, or anti-MAVS antibodies. f SPR analysis of the interactions between MAVS and NS4A. Direct binding was measured by Biacore assays. MAVS was immobilized on a CM5 chip. The analytes consisted of serial dilutions of NS4A proteins ranging between 0 and 2000 nM. The data shown are representative of three independent experiments with similar results
    Figure Legend Snippet: ZIKV NS4A co-localizes and interacts with MAVS. a HeLa cells transfected with plasmids expressing Flag-tagged NS4A, Flag-tagged ZIKV prM or influenza Flag-tagged PB1-F2 were stained with anti-Flag and anti-MAVS antibodies as well as DAPI. Secondary antibodies conjugated to rhodamine and FITC dye were used to visualize the indicated proteins. Images are representative of three independent experiments. 293T cells co-transfected with plasmids encoding Myc-tagged MAVS and Flag-tagged NS4A were used in a co-IP assay to address whether ZIKV NS4A protein physically interacts with MAVS. Cell lysates were precipitated with an anti-Flag antibody ( b ), anti-Myc antibody ( c ), or control mouse IgG, and immunocomplexes were analyzed with the indicated antibodies by western blotting. d 293T cells were transfected with plasmids encoding Flag-tagged NS4A, followed by immunoprecipitation using anti-Flag antibody or control IgG. The immunocomplexes were analyzed with anti-MAVS antibody by Western blotting. e HFF-1 cells were infected with ZIKV at an MOI of 5 followed by immunoprecipitation using anti-MAVS antibody or control mouse IgG. The immunocomplexes that were captured by the protein G Dynabeads were analyzed by Western blotting using anti-NS4A, or anti-MAVS antibodies. f SPR analysis of the interactions between MAVS and NS4A. Direct binding was measured by Biacore assays. MAVS was immobilized on a CM5 chip. The analytes consisted of serial dilutions of NS4A proteins ranging between 0 and 2000 nM. The data shown are representative of three independent experiments with similar results

    Techniques Used: Transfection, Expressing, Staining, Co-Immunoprecipitation Assay, Western Blot, Immunoprecipitation, Infection, SPR Assay, Binding Assay, Chromatin Immunoprecipitation

    6) Product Images from "The effects of DNA formulation and administration route on cancer therapeutic efficacy with xenogenic EGFR DNA vaccine in a lung cancer animal model"

    Article Title: The effects of DNA formulation and administration route on cancer therapeutic efficacy with xenogenic EGFR DNA vaccine in a lung cancer animal model

    Journal: Genetic Vaccines and Therapy

    doi: 10.1186/1479-0556-7-2

    Tumor infiltration of CD4+ and CD8+ T cells . Tumors were excised from mice administrated with Sec-N'-EGFR, or control DNA vector by different delivery methods. Analysis of (A) CD4+ and (B) CD8+ T cells in cryosections of tumors was performed with staining with primary antibody specific for CD4+ and CD8+ cells respectively. Peroxidase-conjugated antibody was used as secondary antibody. Dark spots, peroxidase-stained cells. Similar results were obtained from two more repeated experiments (n = 3 per group).
    Figure Legend Snippet: Tumor infiltration of CD4+ and CD8+ T cells . Tumors were excised from mice administrated with Sec-N'-EGFR, or control DNA vector by different delivery methods. Analysis of (A) CD4+ and (B) CD8+ T cells in cryosections of tumors was performed with staining with primary antibody specific for CD4+ and CD8+ cells respectively. Peroxidase-conjugated antibody was used as secondary antibody. Dark spots, peroxidase-stained cells. Similar results were obtained from two more repeated experiments (n = 3 per group).

    Techniques Used: Mouse Assay, Plasmid Preparation, Staining

    Overexpression of EGFR in LL2 lung cancer cell line . The expression of EGFR in various cell lines was analyzed by Western blotting with monoclonal antibody against EGFR. (B) Flow cytometry analysis of membrane EGFR in LL2 cells. LL2 cells were stained with monoclonal antibody against the extracellular domain of mouse EGFR, followed by FITC-conjugated mouse anti-goat secondary antibody (gray histogram). Normal mouse IgG mAb was used as the negative control (white histogram).
    Figure Legend Snippet: Overexpression of EGFR in LL2 lung cancer cell line . The expression of EGFR in various cell lines was analyzed by Western blotting with monoclonal antibody against EGFR. (B) Flow cytometry analysis of membrane EGFR in LL2 cells. LL2 cells were stained with monoclonal antibody against the extracellular domain of mouse EGFR, followed by FITC-conjugated mouse anti-goat secondary antibody (gray histogram). Normal mouse IgG mAb was used as the negative control (white histogram).

    Techniques Used: Over Expression, Expressing, Western Blot, Flow Cytometry, Staining, Negative Control

    7) Product Images from "Immortalised hippocampal astrocytes from 3xTG-AD mice fail to support BBB integrity in vitro: Role of extracellular vesicles in glial-endothelial communication"

    Article Title: Immortalised hippocampal astrocytes from 3xTG-AD mice fail to support BBB integrity in vitro: Role of extracellular vesicles in glial-endothelial communication

    Journal: bioRxiv

    doi: 10.1101/2020.03.26.009563

    Characterization of extracellular vesicles isolated from WT-iAstro and 3Tg-iAstro ( a ) iAstro derived EVs Nanoparticle Tracking Analysis (NTA) was performed by NanoSight LM10 instrument (Malvern Panalytical). Size distribution of both WT-iAstro and 3Tg-iAstro derived EVs was around 140 nm (n = 3). ( b ) iAstro cell and EV lysates were subjected to electrophoresis, blotted and the membrane was probed with antibodies against EV markers (CD63, HSP70, MFG-E8, syntenin-1). Bands were visualized by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies and chemiluminescence substrate (n = 3). Unprocessed Western blot images are provided in supplements ( c ) Representative electron microphotographs of EVs derived from WT-iAstro and 3Tg-iAstro. Images were acquired using FEI Morgagn 268 transmission electron microscope, iTEM 5.0 software.
    Figure Legend Snippet: Characterization of extracellular vesicles isolated from WT-iAstro and 3Tg-iAstro ( a ) iAstro derived EVs Nanoparticle Tracking Analysis (NTA) was performed by NanoSight LM10 instrument (Malvern Panalytical). Size distribution of both WT-iAstro and 3Tg-iAstro derived EVs was around 140 nm (n = 3). ( b ) iAstro cell and EV lysates were subjected to electrophoresis, blotted and the membrane was probed with antibodies against EV markers (CD63, HSP70, MFG-E8, syntenin-1). Bands were visualized by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies and chemiluminescence substrate (n = 3). Unprocessed Western blot images are provided in supplements ( c ) Representative electron microphotographs of EVs derived from WT-iAstro and 3Tg-iAstro. Images were acquired using FEI Morgagn 268 transmission electron microscope, iTEM 5.0 software.

    Techniques Used: Isolation, Derivative Assay, Electrophoresis, Incubation, Western Blot, Transmission Assay, Microscopy, Software

    8) Product Images from "VX-809 mitigates disease in a mouse model of autosomal dominant polycystic kidney disease bearing the R3277C human mutation"

    Article Title: VX-809 mitigates disease in a mouse model of autosomal dominant polycystic kidney disease bearing the R3277C human mutation

    Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

    doi: 10.1096/fj.202101315R

    Co-localization of CFTR with WGA or Na + /K + ATPase: Images in a normal (wt) mouse, untreated RC/RC mouse, and RC/RC mouse treated with VX-809 (30 mg/kg). Staining for CFTR (red) and WGA or Na + /K + ATPase (green), and the merged image. Yellow denotes co-localization (A,C). Summary of Pearson's correlation coefficient R (B,D). Samples were incubated with primary antibody (CFTR 596 and 488 conjugated WGA or CFTR 596 and Na + /K + ATPase). Anti-mouse-Alexa Fluor 594 secondary or Anti-Rabbit-Alexa Fluor 488 secondary antibodies were used. Scale bar is 10 μm. Dots = number of experimental measurements from three animals, (cortical and medullary regions)
    Figure Legend Snippet: Co-localization of CFTR with WGA or Na + /K + ATPase: Images in a normal (wt) mouse, untreated RC/RC mouse, and RC/RC mouse treated with VX-809 (30 mg/kg). Staining for CFTR (red) and WGA or Na + /K + ATPase (green), and the merged image. Yellow denotes co-localization (A,C). Summary of Pearson's correlation coefficient R (B,D). Samples were incubated with primary antibody (CFTR 596 and 488 conjugated WGA or CFTR 596 and Na + /K + ATPase). Anti-mouse-Alexa Fluor 594 secondary or Anti-Rabbit-Alexa Fluor 488 secondary antibodies were used. Scale bar is 10 μm. Dots = number of experimental measurements from three animals, (cortical and medullary regions)

    Techniques Used: Whole Genome Amplification, Staining, Incubation

    Co-localization of CFTR and WGA or Na + /K + ATPase: Images in a normal (wt) mouse, untreated RC/RC mouse, and RC/RC mouse treated with VX-809 (30 mg/kg) every other day for 60 days beginning at 6 months of age. Staining indicates CFTR (red) and WGA or Na + /K + ATPase (green), and the merged image. Yellow denotes co-localization (A,C). Summary of Pearson's correlation coefficient R (B,D). Samples were incubated with primary antibody (CFTR 596 and 488 conjugated WGA or CFTR 596 and Na + /K + ATPase). Anti-mouse-Alexa Fluor 594 or Anti-Rabbit-Alexa Fluor 488 secondary antibodies were used. Scale bar is 10 μm. Dots = number of experimental measurements from three animals (cortical and medullary regions)
    Figure Legend Snippet: Co-localization of CFTR and WGA or Na + /K + ATPase: Images in a normal (wt) mouse, untreated RC/RC mouse, and RC/RC mouse treated with VX-809 (30 mg/kg) every other day for 60 days beginning at 6 months of age. Staining indicates CFTR (red) and WGA or Na + /K + ATPase (green), and the merged image. Yellow denotes co-localization (A,C). Summary of Pearson's correlation coefficient R (B,D). Samples were incubated with primary antibody (CFTR 596 and 488 conjugated WGA or CFTR 596 and Na + /K + ATPase). Anti-mouse-Alexa Fluor 594 or Anti-Rabbit-Alexa Fluor 488 secondary antibodies were used. Scale bar is 10 μm. Dots = number of experimental measurements from three animals (cortical and medullary regions)

    Techniques Used: Whole Genome Amplification, Staining, Incubation

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    Thermo Fisher hrp conjugated anti mouse igg
    Immunogenicity of triSpike. Sera collected at indicated time points from vaccinated mice or hamsters were analyzed for reactivity with triSpike. (A) Western Blot analysis of pooled sera from immunized mice with or without alum adjuvant. Sera were collected and used at 1/1000 dilution against S-FLAG that was separated under conditions that allowed simultaneous detection of mono-, di-, and trimers of S-protein. A SARS patient serum (SARS), a rabbit serum against S1 and M2 monoclonal antibody against the FLAG peptide was used as control. FLAG-tagged bacterial alkaline phosphatase (BAP-FLAG) was used to assess the presence of antibodies against the FLAG tag. Immune complexes were detected with <t>HRP-conjugated</t> goat anti-mouse, -human or -rabbit <t>IgG</t> polyclonal antibody. Sizes of molecular weight markers are indicated on the right. (B) Reactivity of immune sera from pooled immunized hamster with live BHK-21 cells expressing S-protein at the plasma membrane using FACS analysis. Values were expressed as mean ± standard deviations. (C) Effect of alum adjuvant on longevity of neutralizing response in mouse sera against SARS-CoV (determined on FRhK-4 cells). Values were expressed as mean ± standard deviations. (D) Dose effect of triSpike immunization with alum adjuvant on neutralizing response against SARS-CoV in hamsters (determined on FRhK-4 cells). Values were expressed as mean ± standard deviations. (E) Inhibition of S-protein binding to ACE2 by sera from immunized mice. S-protein coated beads were pre-incubated with sera prior to incubation with soluble ACE2 (sACE2) and detection of the receptor was performed with a polyclonal goat-anti human ACE2 antibody. BAP-FLAG coated beads were used as control.
    Hrp Conjugated Anti Mouse Igg, supplied by Thermo Fisher, 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/hrp conjugated anti mouse igg/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Thermo Fisher horseradish peroxidase conjugated secondary antibodies anti mouse
    Electron micrographs of negative staining and immunogold labeling of VLPs and ZIKV. Purified VLPs and ZIKV were examined by transmission electron microscopy (TEM). Panels A and B show uranyl acetate negatively stained VLPs which exhibit particle sizes from 50nm to 65nm in diameter (mean 60nm) and a structure that resembles the morphology and surface appearance of wild type Zika virus which is shown in Panel E. Immunogold labeling with two antibodies, <t>mouse</t> <t>anti-E</t> protein MAb 4G2 and a human serum from a Zika patient served as primary and counterstained with anti-mouse or anti-human <t>secondary</t> antibody <t>conjugated</t> with gold beads of 6 nm and 10nm in diameter, respectively. Both antibodies, 4G2 panel C and human serum panel D, bind to the particle surfaces as revealed by the presence of gold beads. Panel F shows wild-type ZIKV probed with the 4G2 antibody, which also binds the virus surface as revealed by the detection of gold beads; in this case goat anti-mouse secondary antibody conjugated with 10 nm gold bead was applied. These studies demonstrate that a specific anti-E MAb and an anti-Zika polyclonal antibody reacts with the VLP surfaces indicating that the major Zika surface antigen, the E glycoprotein, is indeed displayed on the VLP surfaces.
    Horseradish Peroxidase Conjugated Secondary Antibodies Anti Mouse, supplied by Thermo Fisher, 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/horseradish peroxidase conjugated secondary antibodies anti mouse/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    horseradish peroxidase conjugated secondary antibodies anti mouse - by Bioz Stars, 2022-09
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    94
    Thermo Fisher anti myc hrp
    The 69-kDa Ku86v in MM cell lines is present in the cytosolic, nuclear and membrane fractions and binds DNA . Having determined that MM cell lines constitutively express 69-kDa Ku86v, we next studied the subcellular location of Ku86v. Moreover, since CD40 triggering induces expression of 86-kDa <t>Ku86</t> on the cell membrane of MM cells, we also investigated whether CD40 triggering affected the subcellular location Ku86v. Fresh whole cell (W), cytosolic (C), nuclear (N) and membrane (M) cell lysate fractions were obtained from sCD40L-triggered (5.0 μg/mL for 4 hrs) and resting RPMI 8226 MM cells, and subjected to normal SDS-PAGE (A). Full-length and variant forms of Ku86 were detected by western immunoblotting using S10B1 anti-Ku86 mAb. Membranes were stripped and re-probed using anti-actin mAb (control) to confirm equal protein loading. Relative expression of 69-kDa Ku86v (normalized to weakest band) was determined using image densitometry and expressed in bar chart format. The presence of Ku86/Ku86v protein-DNA complexes were next detected using EMSA (B and C). Cytosolic (C), nuclear (N) or membrane (M) protein extracts (4.0 μg/sample) were first obtained from CESS (negative control), CD40-triggered (5.0 μg/mL sCD40L for 4 hrs) or non-CD40-triggered RPMI 8226 MM cell lines. Non-competitive binding of Ku86/Ku86v to DNA (B) was detected using a specific biotin end-labeled DNA probe (20.0 fmol/sample), native PAGE, and <t>hrp-conjugated</t> streptavidin chemiluminescence imaging. Full-length Ku86-DNA complexes are found at position I, whereas DNA complexes with the truncation variant of Ku86 are found at position II. Relative expression of 69-kDa Ku86v-DNA complexes (normalized to weakest band) was determined using image densitometry. To confirm the specificity of DNA binding, a competitive assay (C), in which a variable amount of cell lysate (up to 5.0 μg/sample) was mixed with a fixed amount (4.0 pmol/sample) of non-biotin-labeled DNA, was also performed in the same fashion. All experiments were repeated with the SGH-MM5 MM cell line (data not shown), showed similar results, and performed in triplicate.
    Anti Myc Hrp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher peroxidase conjugated goat anti human igg
    <t>Immunoblots</t> to determine reactivities of <t>IgG</t> antibodies to recombinant antigens and control proteins. Nonrecombinant control proteins MBP (lanes 1) and GST (lanes 2) and recombinant antigens rP22 (lanes 3), rP25 (lanes 4), rP29 (lanes 5), and rP35 (lanes 6) were electrophoresed in a sodium dodecyl sulfate–10% polyacrylamide gel and transferred to nitrocellulose paper. Duplicate blots were reacted with a pool of either group I (A) or group II (B) sera. Numbers on the left are molecular masses, in kilodaltons.
    Peroxidase Conjugated Goat Anti Human Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Immunogenicity of triSpike. Sera collected at indicated time points from vaccinated mice or hamsters were analyzed for reactivity with triSpike. (A) Western Blot analysis of pooled sera from immunized mice with or without alum adjuvant. Sera were collected and used at 1/1000 dilution against S-FLAG that was separated under conditions that allowed simultaneous detection of mono-, di-, and trimers of S-protein. A SARS patient serum (SARS), a rabbit serum against S1 and M2 monoclonal antibody against the FLAG peptide was used as control. FLAG-tagged bacterial alkaline phosphatase (BAP-FLAG) was used to assess the presence of antibodies against the FLAG tag. Immune complexes were detected with HRP-conjugated goat anti-mouse, -human or -rabbit IgG polyclonal antibody. Sizes of molecular weight markers are indicated on the right. (B) Reactivity of immune sera from pooled immunized hamster with live BHK-21 cells expressing S-protein at the plasma membrane using FACS analysis. Values were expressed as mean ± standard deviations. (C) Effect of alum adjuvant on longevity of neutralizing response in mouse sera against SARS-CoV (determined on FRhK-4 cells). Values were expressed as mean ± standard deviations. (D) Dose effect of triSpike immunization with alum adjuvant on neutralizing response against SARS-CoV in hamsters (determined on FRhK-4 cells). Values were expressed as mean ± standard deviations. (E) Inhibition of S-protein binding to ACE2 by sera from immunized mice. S-protein coated beads were pre-incubated with sera prior to incubation with soluble ACE2 (sACE2) and detection of the receptor was performed with a polyclonal goat-anti human ACE2 antibody. BAP-FLAG coated beads were used as control.

    Journal: Vaccine

    Article Title: Antibodies against trimeric S glycoprotein protect hamsters against SARS-CoV challenge despite their capacity to mediate FcγRII-dependent entry into B cells in vitro

    doi: 10.1016/j.vaccine.2006.08.011

    Figure Lengend Snippet: Immunogenicity of triSpike. Sera collected at indicated time points from vaccinated mice or hamsters were analyzed for reactivity with triSpike. (A) Western Blot analysis of pooled sera from immunized mice with or without alum adjuvant. Sera were collected and used at 1/1000 dilution against S-FLAG that was separated under conditions that allowed simultaneous detection of mono-, di-, and trimers of S-protein. A SARS patient serum (SARS), a rabbit serum against S1 and M2 monoclonal antibody against the FLAG peptide was used as control. FLAG-tagged bacterial alkaline phosphatase (BAP-FLAG) was used to assess the presence of antibodies against the FLAG tag. Immune complexes were detected with HRP-conjugated goat anti-mouse, -human or -rabbit IgG polyclonal antibody. Sizes of molecular weight markers are indicated on the right. (B) Reactivity of immune sera from pooled immunized hamster with live BHK-21 cells expressing S-protein at the plasma membrane using FACS analysis. Values were expressed as mean ± standard deviations. (C) Effect of alum adjuvant on longevity of neutralizing response in mouse sera against SARS-CoV (determined on FRhK-4 cells). Values were expressed as mean ± standard deviations. (D) Dose effect of triSpike immunization with alum adjuvant on neutralizing response against SARS-CoV in hamsters (determined on FRhK-4 cells). Values were expressed as mean ± standard deviations. (E) Inhibition of S-protein binding to ACE2 by sera from immunized mice. S-protein coated beads were pre-incubated with sera prior to incubation with soluble ACE2 (sACE2) and detection of the receptor was performed with a polyclonal goat-anti human ACE2 antibody. BAP-FLAG coated beads were used as control.

    Article Snippet: HRP-conjugated anti-mouse IgG (H + L) or IgA (1/1000) (Zymed) were used to detect the existence of IgG or IgA antibody against triSpike in immunized mice fecal and nasal lavage sample.

    Techniques: Mouse Assay, Western Blot, FLAG-tag, Molecular Weight, Expressing, FACS, Inhibition, Protein Binding, Incubation

    Induction of mucosal immune response in triSpike plus alum vaccinated mice. (A) Antibodies prepared from fecal samples collected at day 44 post-immunization were reacted against S-protein as described in Fig. 2 A. Immune complexes were detected with HRP-conjugated goat anti-mouse IgG or IgA polyclonal antibody. (B) Similar as (A) except that Western Blot analysis was performed with pooled nasal lavage samples collected at day 65. (C). Antibodies prepared from fecal samples of vaccinated mice were analyzed for neutralizing activity against SARS-CoV infection on FRhK-4 cells. Weak neutralizing activity was detected after the third immunization only. Asterisk (**) indicates the value of p

    Journal: Vaccine

    Article Title: Antibodies against trimeric S glycoprotein protect hamsters against SARS-CoV challenge despite their capacity to mediate FcγRII-dependent entry into B cells in vitro

    doi: 10.1016/j.vaccine.2006.08.011

    Figure Lengend Snippet: Induction of mucosal immune response in triSpike plus alum vaccinated mice. (A) Antibodies prepared from fecal samples collected at day 44 post-immunization were reacted against S-protein as described in Fig. 2 A. Immune complexes were detected with HRP-conjugated goat anti-mouse IgG or IgA polyclonal antibody. (B) Similar as (A) except that Western Blot analysis was performed with pooled nasal lavage samples collected at day 65. (C). Antibodies prepared from fecal samples of vaccinated mice were analyzed for neutralizing activity against SARS-CoV infection on FRhK-4 cells. Weak neutralizing activity was detected after the third immunization only. Asterisk (**) indicates the value of p

    Article Snippet: HRP-conjugated anti-mouse IgG (H + L) or IgA (1/1000) (Zymed) were used to detect the existence of IgG or IgA antibody against triSpike in immunized mice fecal and nasal lavage sample.

    Techniques: Mouse Assay, Western Blot, Activity Assay, Infection

    Electron micrographs of negative staining and immunogold labeling of VLPs and ZIKV. Purified VLPs and ZIKV were examined by transmission electron microscopy (TEM). Panels A and B show uranyl acetate negatively stained VLPs which exhibit particle sizes from 50nm to 65nm in diameter (mean 60nm) and a structure that resembles the morphology and surface appearance of wild type Zika virus which is shown in Panel E. Immunogold labeling with two antibodies, mouse anti-E protein MAb 4G2 and a human serum from a Zika patient served as primary and counterstained with anti-mouse or anti-human secondary antibody conjugated with gold beads of 6 nm and 10nm in diameter, respectively. Both antibodies, 4G2 panel C and human serum panel D, bind to the particle surfaces as revealed by the presence of gold beads. Panel F shows wild-type ZIKV probed with the 4G2 antibody, which also binds the virus surface as revealed by the detection of gold beads; in this case goat anti-mouse secondary antibody conjugated with 10 nm gold bead was applied. These studies demonstrate that a specific anti-E MAb and an anti-Zika polyclonal antibody reacts with the VLP surfaces indicating that the major Zika surface antigen, the E glycoprotein, is indeed displayed on the VLP surfaces.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Zika virus-like particle (VLP) based vaccine

    doi: 10.1371/journal.pntd.0005608

    Figure Lengend Snippet: Electron micrographs of negative staining and immunogold labeling of VLPs and ZIKV. Purified VLPs and ZIKV were examined by transmission electron microscopy (TEM). Panels A and B show uranyl acetate negatively stained VLPs which exhibit particle sizes from 50nm to 65nm in diameter (mean 60nm) and a structure that resembles the morphology and surface appearance of wild type Zika virus which is shown in Panel E. Immunogold labeling with two antibodies, mouse anti-E protein MAb 4G2 and a human serum from a Zika patient served as primary and counterstained with anti-mouse or anti-human secondary antibody conjugated with gold beads of 6 nm and 10nm in diameter, respectively. Both antibodies, 4G2 panel C and human serum panel D, bind to the particle surfaces as revealed by the presence of gold beads. Panel F shows wild-type ZIKV probed with the 4G2 antibody, which also binds the virus surface as revealed by the detection of gold beads; in this case goat anti-mouse secondary antibody conjugated with 10 nm gold bead was applied. These studies demonstrate that a specific anti-E MAb and an anti-Zika polyclonal antibody reacts with the VLP surfaces indicating that the major Zika surface antigen, the E glycoprotein, is indeed displayed on the VLP surfaces.

    Article Snippet: Horseradish peroxidase conjugated secondary antibodies anti-mouse and anti-rabbit were purchased from Pierce Thermo Fisher, MA (#31430 and# 31460 respectively).

    Techniques: Negative Staining, Labeling, Purification, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Staining

    The 69-kDa Ku86v in MM cell lines is present in the cytosolic, nuclear and membrane fractions and binds DNA . Having determined that MM cell lines constitutively express 69-kDa Ku86v, we next studied the subcellular location of Ku86v. Moreover, since CD40 triggering induces expression of 86-kDa Ku86 on the cell membrane of MM cells, we also investigated whether CD40 triggering affected the subcellular location Ku86v. Fresh whole cell (W), cytosolic (C), nuclear (N) and membrane (M) cell lysate fractions were obtained from sCD40L-triggered (5.0 μg/mL for 4 hrs) and resting RPMI 8226 MM cells, and subjected to normal SDS-PAGE (A). Full-length and variant forms of Ku86 were detected by western immunoblotting using S10B1 anti-Ku86 mAb. Membranes were stripped and re-probed using anti-actin mAb (control) to confirm equal protein loading. Relative expression of 69-kDa Ku86v (normalized to weakest band) was determined using image densitometry and expressed in bar chart format. The presence of Ku86/Ku86v protein-DNA complexes were next detected using EMSA (B and C). Cytosolic (C), nuclear (N) or membrane (M) protein extracts (4.0 μg/sample) were first obtained from CESS (negative control), CD40-triggered (5.0 μg/mL sCD40L for 4 hrs) or non-CD40-triggered RPMI 8226 MM cell lines. Non-competitive binding of Ku86/Ku86v to DNA (B) was detected using a specific biotin end-labeled DNA probe (20.0 fmol/sample), native PAGE, and hrp-conjugated streptavidin chemiluminescence imaging. Full-length Ku86-DNA complexes are found at position I, whereas DNA complexes with the truncation variant of Ku86 are found at position II. Relative expression of 69-kDa Ku86v-DNA complexes (normalized to weakest band) was determined using image densitometry. To confirm the specificity of DNA binding, a competitive assay (C), in which a variable amount of cell lysate (up to 5.0 μg/sample) was mixed with a fixed amount (4.0 pmol/sample) of non-biotin-labeled DNA, was also performed in the same fashion. All experiments were repeated with the SGH-MM5 MM cell line (data not shown), showed similar results, and performed in triplicate.

    Journal: Cancer Cell International

    Article Title: Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells

    doi: 10.1186/1475-2867-8-4

    Figure Lengend Snippet: The 69-kDa Ku86v in MM cell lines is present in the cytosolic, nuclear and membrane fractions and binds DNA . Having determined that MM cell lines constitutively express 69-kDa Ku86v, we next studied the subcellular location of Ku86v. Moreover, since CD40 triggering induces expression of 86-kDa Ku86 on the cell membrane of MM cells, we also investigated whether CD40 triggering affected the subcellular location Ku86v. Fresh whole cell (W), cytosolic (C), nuclear (N) and membrane (M) cell lysate fractions were obtained from sCD40L-triggered (5.0 μg/mL for 4 hrs) and resting RPMI 8226 MM cells, and subjected to normal SDS-PAGE (A). Full-length and variant forms of Ku86 were detected by western immunoblotting using S10B1 anti-Ku86 mAb. Membranes were stripped and re-probed using anti-actin mAb (control) to confirm equal protein loading. Relative expression of 69-kDa Ku86v (normalized to weakest band) was determined using image densitometry and expressed in bar chart format. The presence of Ku86/Ku86v protein-DNA complexes were next detected using EMSA (B and C). Cytosolic (C), nuclear (N) or membrane (M) protein extracts (4.0 μg/sample) were first obtained from CESS (negative control), CD40-triggered (5.0 μg/mL sCD40L for 4 hrs) or non-CD40-triggered RPMI 8226 MM cell lines. Non-competitive binding of Ku86/Ku86v to DNA (B) was detected using a specific biotin end-labeled DNA probe (20.0 fmol/sample), native PAGE, and hrp-conjugated streptavidin chemiluminescence imaging. Full-length Ku86-DNA complexes are found at position I, whereas DNA complexes with the truncation variant of Ku86 are found at position II. Relative expression of 69-kDa Ku86v-DNA complexes (normalized to weakest band) was determined using image densitometry. To confirm the specificity of DNA binding, a competitive assay (C), in which a variable amount of cell lysate (up to 5.0 μg/sample) was mixed with a fixed amount (4.0 pmol/sample) of non-biotin-labeled DNA, was also performed in the same fashion. All experiments were repeated with the SGH-MM5 MM cell line (data not shown), showed similar results, and performed in triplicate.

    Article Snippet: Finally, the Ku86 recombinant proteins were purified using ProBond™ Nickel-Chelating Resin column (Invitrogen) and detected using western Immunblotting with anti-myc-HRP (Invitrogen) and/or anti-Ku86 antibody (Neomarker).

    Techniques: Expressing, SDS Page, Variant Assay, Western Blot, Negative Control, Binding Assay, Labeling, Clear Native PAGE, Imaging

    Immunoblots to determine reactivities of IgG antibodies to recombinant antigens and control proteins. Nonrecombinant control proteins MBP (lanes 1) and GST (lanes 2) and recombinant antigens rP22 (lanes 3), rP25 (lanes 4), rP29 (lanes 5), and rP35 (lanes 6) were electrophoresed in a sodium dodecyl sulfate–10% polyacrylamide gel and transferred to nitrocellulose paper. Duplicate blots were reacted with a pool of either group I (A) or group II (B) sera. Numbers on the left are molecular masses, in kilodaltons.

    Journal: Clinical and Diagnostic Laboratory Immunology

    Article Title: Serodiagnosis of Recently Acquired Toxoplasma gondii Infection Using an Enzyme-Linked Immunosorbent Assay with a Combination of Recombinant Antigens

    doi:

    Figure Lengend Snippet: Immunoblots to determine reactivities of IgG antibodies to recombinant antigens and control proteins. Nonrecombinant control proteins MBP (lanes 1) and GST (lanes 2) and recombinant antigens rP22 (lanes 3), rP25 (lanes 4), rP29 (lanes 5), and rP35 (lanes 6) were electrophoresed in a sodium dodecyl sulfate–10% polyacrylamide gel and transferred to nitrocellulose paper. Duplicate blots were reacted with a pool of either group I (A) or group II (B) sera. Numbers on the left are molecular masses, in kilodaltons.

    Article Snippet: Thereafter, the immunoblots were incubated with horseradish peroxidase-conjugated goat anti-human IgG (Caltag Laboratories, Burlingame, Calif.) at an optimal dilution of 1:8,000 in phosphate-buffered saline (PBS) with 3% bovine serum albumin (BSA) at room temperature for 1 h. After being washed with PBS, the membranes were incubated with 3,3′-diaminobenzidine tetrahydrochloride (Sigma Chemicals) at a concentration of 0.1 mg/ml in PBS.

    Techniques: Western Blot, Recombinant