hrp-conjugated goat anti-rabbit igg Search Results


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  • 99
    Bio-Rad hrp conjugated goat anti rabbit igg
    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with <t>HRP-conjugated</t> anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat <t>IgG</t> and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 526 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti rabbit igg/product/Bio-Rad
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    99
    Bio-Rad hrp conjugated goat anti rabbit
    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with <t>HRP-conjugated</t> anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat <t>IgG</t> and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.
    Hrp Conjugated Goat Anti Rabbit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 367 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti rabbit/product/Bio-Rad
    Average 99 stars, based on 367 article reviews
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    99
    Bio-Rad goat anti rabbit igg hrp conjugated
    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with <t>HRP-conjugated</t> anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat <t>IgG</t> and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.
    Goat Anti Rabbit Igg Hrp Conjugated, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit igg hrp conjugated/product/Bio-Rad
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    99
    Bio-Rad goat anti rabbit hrp conjugated
    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with <t>HRP-conjugated</t> anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat <t>IgG</t> and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.
    Goat Anti Rabbit Hrp Conjugated, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 172 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti rabbit hrp conjugated/product/Bio-Rad
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    96
    SouthernBiotech hrp conjugated goat anti rabbit igg
    Highly specific binding of plant-derived E16 to WNV DIII antigen. WNV DIII or DENV-2 DIII antigen produced in E. coli was immobilized on an ELISA plate and incubated with increasing concentrations of lettuce or N. benthamiana -derived E16 mAb (L-E16 or B-E16), mammalian cell-derived E16 (M-E16, positive control), or a negative control generic human <t>IgG</t> (NC). A <t>HRP-conjugated</t> anti-human IgG was used to detect mAbs bound to DIII antigens. Mean ± SD of OD 450 from three independent experiments are presented.
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 96/100, based on 172 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti rabbit igg/product/SouthernBiotech
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    92
    Proteintech hrp conjugated goat anti rabbit igg
    Highly specific binding of plant-derived E16 to WNV DIII antigen. WNV DIII or DENV-2 DIII antigen produced in E. coli was immobilized on an ELISA plate and incubated with increasing concentrations of lettuce or N. benthamiana -derived E16 mAb (L-E16 or B-E16), mammalian cell-derived E16 (M-E16, positive control), or a negative control generic human <t>IgG</t> (NC). A <t>HRP-conjugated</t> anti-human IgG was used to detect mAbs bound to DIII antigens. Mean ± SD of OD 450 from three independent experiments are presented.
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Stressgen Biotechnologies hrp conjugated goat anti rabbit igg
    Highly specific binding of plant-derived E16 to WNV DIII antigen. WNV DIII or DENV-2 DIII antigen produced in E. coli was immobilized on an ELISA plate and incubated with increasing concentrations of lettuce or N. benthamiana -derived E16 mAb (L-E16 or B-E16), mammalian cell-derived E16 (M-E16, positive control), or a negative control generic human <t>IgG</t> (NC). A <t>HRP-conjugated</t> anti-human IgG was used to detect mAbs bound to DIII antigens. Mean ± SD of OD 450 from three independent experiments are presented.
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by Stressgen Biotechnologies, 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/hrp conjugated goat anti rabbit igg/product/Stressgen Biotechnologies
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    94
    Thermo Fisher hrp conjugated goat anti rabbit igg
    Binding activity of human dsNKG2D–IL-15 with MICA. ( a ) The human dsNKG2D–IL-15 protein was identified by the NKG2D antibody or IL-15 antibody through a western blot assay. ( b ) Plates coated with human dsNKG2D–IL-15, human NKG2D–Ig proteins, and IL-15 were bound by a human NKG2D antibody or an IL-15 antibody and were detected by ELISA. ( c ) Wells were coated with a soluble MICA protein (1 μg) or soluble RAE-1ε (1 μg). Human dsNKG2D–IL-15 was added and incubated at 37°C for 1 h. The NKG2D mAb, NKG2D pAb, or IL-15 pAb and <t>HRP-conjugated</t> secondary antibodies were added sequentially to the wells. The OD 450 value of each well was read after the addition of the substrate and stop solution. ( d ) All tumor cells were incubated with various concentrations of human dsNKG2D–IL-15, stained with a fluorescent IL-15 mAb and detected by flow cytometry. Gray area: isotype control; dotted line: IL-15 (5 μg mL −1 ) incubation and IL-15 mAb staining; and dashed line: human dsNKG2D–IL-15 incubation and IL-15 mAb staining. ( e ) B16-MICA cells were incubated with human or mouse dsNKG2D–IL-15 and detected with an IL-15 pAb, NKG2D mAb, or NKG2D pAb with or without MICA mAb neutralization. ( f ) B16BL6–MICA cells were co-incubated with serial concentrations of the human dsNKG2D–IL-15, mouse dsNKG2D–IL-15 or human NKG2D–Ig proteins (5 μg mL −1 ), stained with goat anti-human <t>IgG</t> antibody, and detected by flow cytometry. * P
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1334 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Jackson Immuno goat anti rabbit igg conjugated hrp
    Binding activity of human dsNKG2D–IL-15 with MICA. ( a ) The human dsNKG2D–IL-15 protein was identified by the NKG2D antibody or IL-15 antibody through a western blot assay. ( b ) Plates coated with human dsNKG2D–IL-15, human NKG2D–Ig proteins, and IL-15 were bound by a human NKG2D antibody or an IL-15 antibody and were detected by ELISA. ( c ) Wells were coated with a soluble MICA protein (1 μg) or soluble RAE-1ε (1 μg). Human dsNKG2D–IL-15 was added and incubated at 37°C for 1 h. The NKG2D mAb, NKG2D pAb, or IL-15 pAb and <t>HRP-conjugated</t> secondary antibodies were added sequentially to the wells. The OD 450 value of each well was read after the addition of the substrate and stop solution. ( d ) All tumor cells were incubated with various concentrations of human dsNKG2D–IL-15, stained with a fluorescent IL-15 mAb and detected by flow cytometry. Gray area: isotype control; dotted line: IL-15 (5 μg mL −1 ) incubation and IL-15 mAb staining; and dashed line: human dsNKG2D–IL-15 incubation and IL-15 mAb staining. ( e ) B16-MICA cells were incubated with human or mouse dsNKG2D–IL-15 and detected with an IL-15 pAb, NKG2D mAb, or NKG2D pAb with or without MICA mAb neutralization. ( f ) B16BL6–MICA cells were co-incubated with serial concentrations of the human dsNKG2D–IL-15, mouse dsNKG2D–IL-15 or human NKG2D–Ig proteins (5 μg mL −1 ), stained with goat anti-human <t>IgG</t> antibody, and detected by flow cytometry. * P
    Goat Anti Rabbit Igg Conjugated Hrp, supplied by Jackson Immuno, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam hrp conjugated goat anti rabbit igg
    Binding ELISA demonstrating the autoantibody binding to Met5A mesothelial cells due to the presence of MCAA in serum from human patients exposed to LA fibers. Sera were pooled from MCAA positive samples (MCAA+) and MCAA-positive samples cleared of all <t>IgG</t> (Cleared). Human mesothelial cells were plated in 96-well plates and then stained for MCAA binding using the pooled serum as the primary antibody and anti-human IgG - <t>HRP</t> for the secondary antibody. N= 3 samples per treatment group. Data are mean absorbance at 450 nm. *=p
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 461 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Beijing CWBio hrp conjugated goat anti rabbit igg
    Binding ELISA demonstrating the autoantibody binding to Met5A mesothelial cells due to the presence of MCAA in serum from human patients exposed to LA fibers. Sera were pooled from MCAA positive samples (MCAA+) and MCAA-positive samples cleared of all <t>IgG</t> (Cleared). Human mesothelial cells were plated in 96-well plates and then stained for MCAA binding using the pooled serum as the primary antibody and anti-human IgG - <t>HRP</t> for the secondary antibody. N= 3 samples per treatment group. Data are mean absorbance at 450 nm. *=p
    Hrp Conjugated Goat Anti Rabbit Igg, supplied by Beijing CWBio, used in various techniques. Bioz Stars score: 92/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti rabbit igg/product/Beijing CWBio
    Average 92 stars, based on 57 article reviews
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    Image Search Results


    SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with HRP-conjugated anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat IgG and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.

    Journal: The Journal of Biological Chemistry

    Article Title: New Insights into the DT40 B Cell Receptor Cluster Using a Proteomic Proximity Labeling Assay *

    doi: 10.1074/jbc.M113.529578

    Figure Lengend Snippet: SPPLAT analysis of the BCR-induced clusters in DT40 cells. A , co-localization of BCR and deposited biotin. Cells were preincubated with HRP-conjugated anti-chicken IgM and tyramide biotinylated. The distribution of BCR and deposited biotin was detected using FITC-conjugated anti-goat IgG and Alexa 568-labeled avidin as described under “Experimental Procedures.” Three representative images are shown with corresponding images rendered as pseudo-three-dimensional. Bar = 5 μm. B , tyramide-biotinylated cells were stripped of biotin by treatment with impermeant reducing agent as described under “Experimental Procedures.” Cells were stained with FITC-conjugated anti-goat IgG to detect BCR, and with Texas Red-conjugated avidin to detect remaining biotin. Bar = 5 μm.

    Article Snippet: About 5 × 108 cells were incubated with end over end rotation for 2 h with 20 μg/ml of HRP-conjugated goat anti-chicken IgM (Bethyl Ltd.), or 20 μg/ml of HRP-conjugated goat anti-rabbit IgG (Bio-Rad) in 20 ml of PBS with 10% (v/v) goat serum (Ultraclone Ltd.) at 4 °C.

    Techniques: Labeling, Avidin-Biotin Assay, Staining

    Immunoisolation of E1-G CT containing membranes. BHK-E1-G CT cells were homogenized and separated into total homogenate (TH), postnuclear supernatant (PNS), nuclear pellet (NP), smooth membranes (SM), rough membranes (RM), and immunoisolate fractions (bound) prepared using P5D4-coated magnetic beads. Equivalent volumes normalized to starting material for each fraction were electrophoresed through 10% polyacrylamide gels followed by transfer to PVDF membranes. E1-G CT was detected by probing membranes with rabbit antibody directed against the CT domain of VSV G followed by goat anti-rabbit IgG-HRP and ECL detection. The smooth membrane fraction was used as the starting material for immunoisolation. Most of the E1-G CT was recovered in the bound fraction; however, a significant portion did not bind to the beads (unbound).

    Journal: Molecular Biology of the Cell

    Article Title: Immunoisolation and Characterization of a Subdomain of the Endoplasmic Reticulum That Concentrates Proteins Involved in COPII Vesicle Biogenesis

    doi:

    Figure Lengend Snippet: Immunoisolation of E1-G CT containing membranes. BHK-E1-G CT cells were homogenized and separated into total homogenate (TH), postnuclear supernatant (PNS), nuclear pellet (NP), smooth membranes (SM), rough membranes (RM), and immunoisolate fractions (bound) prepared using P5D4-coated magnetic beads. Equivalent volumes normalized to starting material for each fraction were electrophoresed through 10% polyacrylamide gels followed by transfer to PVDF membranes. E1-G CT was detected by probing membranes with rabbit antibody directed against the CT domain of VSV G followed by goat anti-rabbit IgG-HRP and ECL detection. The smooth membrane fraction was used as the starting material for immunoisolation. Most of the E1-G CT was recovered in the bound fraction; however, a significant portion did not bind to the beads (unbound).

    Article Snippet: Secondary antibodies, goat anti-mouse IgG, and goat anti-rabbit IgG conjugated to HRP were obtained from Bio-Rad (Richmond, CA).

    Techniques: Magnetic Beads

    Immunoblotting of P5D4-immunoisolated membranes with anti-ER, Golgi and ERGIC markers. BHK-E1-G CT cells were homogenized and separated into postnuclear supernatant (PNS), rough microsomes (RM), smooth membranes (SM), P5D4-immunoisolates (P5D4), and immunoisolates obtained using B8G65, a mAb of the same isotype as P5D4, that recognizes the ectodomain of VSV-G. Immunoisolates of PNS, RM, and SM (15 μg each) and P5D4 and BW8G65 (3 μg each) were separated on 10% gels before transfer to PVDF. Blots were probed with rabbit anti-VSV G CT serum (to detect E1-G CT ) and various antibodies to marker proteins for ER, Golgi, and ERGIC followed by goat anti-rabbit and anti-mouse IgG conjugated to HRP. Blots were then subjected to ECL development. (A) Soluble RER proteins and the ERGIC/Golgi marker β-COP are largely depleted or absent from the P5D4 immunoisolates. In contrast, E1-G CT and Sec13p are enriched. (B) The ERGIC marker p58 is highly enriched in the E1-G CT –containing membranes.

    Journal: Molecular Biology of the Cell

    Article Title: Immunoisolation and Characterization of a Subdomain of the Endoplasmic Reticulum That Concentrates Proteins Involved in COPII Vesicle Biogenesis

    doi:

    Figure Lengend Snippet: Immunoblotting of P5D4-immunoisolated membranes with anti-ER, Golgi and ERGIC markers. BHK-E1-G CT cells were homogenized and separated into postnuclear supernatant (PNS), rough microsomes (RM), smooth membranes (SM), P5D4-immunoisolates (P5D4), and immunoisolates obtained using B8G65, a mAb of the same isotype as P5D4, that recognizes the ectodomain of VSV-G. Immunoisolates of PNS, RM, and SM (15 μg each) and P5D4 and BW8G65 (3 μg each) were separated on 10% gels before transfer to PVDF. Blots were probed with rabbit anti-VSV G CT serum (to detect E1-G CT ) and various antibodies to marker proteins for ER, Golgi, and ERGIC followed by goat anti-rabbit and anti-mouse IgG conjugated to HRP. Blots were then subjected to ECL development. (A) Soluble RER proteins and the ERGIC/Golgi marker β-COP are largely depleted or absent from the P5D4 immunoisolates. In contrast, E1-G CT and Sec13p are enriched. (B) The ERGIC marker p58 is highly enriched in the E1-G CT –containing membranes.

    Article Snippet: Secondary antibodies, goat anti-mouse IgG, and goat anti-rabbit IgG conjugated to HRP were obtained from Bio-Rad (Richmond, CA).

    Techniques: Marker

    A : immunofluorescent study shows hAT 1 R is downregulated in cells exposed to 25 mM glucose. Immunofluorescent staining used primary rabbit anti-hAT 1 R IgG followed by secondary anti-rabbit IgG conjugated with Alexa Fluor 488. Nuclei were stained with 4′,6-diamidino-2-phenylindole

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Functional role of sodium glucose transporter in high glucose-mediated angiotensin type 1 receptor downregulation in human proximal tubule cells

    doi: 10.1152/ajprenal.00651.2011

    Figure Lengend Snippet: A : immunofluorescent study shows hAT 1 R is downregulated in cells exposed to 25 mM glucose. Immunofluorescent staining used primary rabbit anti-hAT 1 R IgG followed by secondary anti-rabbit IgG conjugated with Alexa Fluor 488. Nuclei were stained with 4′,6-diamidino-2-phenylindole

    Article Snippet: The AT1 R (N-10) antibody was from Santa Cruz Biotechnology (Santa Cruz, CA), goat-anti rabbit IgG-horseradish peroxidase conjugate was from Bio-Rad (Hercules, CA), Alexa Fluor goat anti-rabbit IgG, and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were from Invitrogen.

    Techniques: HAT Assay, Staining

    hAT 1 R-specific immunofluorescent studies in cells exposed to NG ( A ), HG ( B ), HG with 0.5 mM Pzin ( C ), and HG with 0.15 mM Ptin ( D ). Immunofluorescent staining was done using primary polyclonal anti-hAT 1 R IgG followed by secondary IgG conjugated with Alexa

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Functional role of sodium glucose transporter in high glucose-mediated angiotensin type 1 receptor downregulation in human proximal tubule cells

    doi: 10.1152/ajprenal.00651.2011

    Figure Lengend Snippet: hAT 1 R-specific immunofluorescent studies in cells exposed to NG ( A ), HG ( B ), HG with 0.5 mM Pzin ( C ), and HG with 0.15 mM Ptin ( D ). Immunofluorescent staining was done using primary polyclonal anti-hAT 1 R IgG followed by secondary IgG conjugated with Alexa

    Article Snippet: The AT1 R (N-10) antibody was from Santa Cruz Biotechnology (Santa Cruz, CA), goat-anti rabbit IgG-horseradish peroxidase conjugate was from Bio-Rad (Hercules, CA), Alexa Fluor goat anti-rabbit IgG, and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were from Invitrogen.

    Techniques: HAT Assay, Staining

    Highly specific binding of plant-derived E16 to WNV DIII antigen. WNV DIII or DENV-2 DIII antigen produced in E. coli was immobilized on an ELISA plate and incubated with increasing concentrations of lettuce or N. benthamiana -derived E16 mAb (L-E16 or B-E16), mammalian cell-derived E16 (M-E16, positive control), or a negative control generic human IgG (NC). A HRP-conjugated anti-human IgG was used to detect mAbs bound to DIII antigens. Mean ± SD of OD 450 from three independent experiments are presented.

    Journal: Journal of Biomedicine and Biotechnology

    Article Title: A Novel System for Rapid and Cost-Effective Production of Detection and Diagnostic Reagents of West Nile Virus in Plants

    doi: 10.1155/2012/106783

    Figure Lengend Snippet: Highly specific binding of plant-derived E16 to WNV DIII antigen. WNV DIII or DENV-2 DIII antigen produced in E. coli was immobilized on an ELISA plate and incubated with increasing concentrations of lettuce or N. benthamiana -derived E16 mAb (L-E16 or B-E16), mammalian cell-derived E16 (M-E16, positive control), or a negative control generic human IgG (NC). A HRP-conjugated anti-human IgG was used to detect mAbs bound to DIII antigens. Mean ± SD of OD 450 from three independent experiments are presented.

    Article Snippet: For DIII antigen, a rabbit anti-WNV DIII polyclonal antibody and an HRP-conjugated goat anti-rabbit IgG (Southern Biotech, Birmingham, Ala, USA) were used for the analysis.

    Techniques: Binding Assay, Derivative Assay, Produced, Enzyme-linked Immunosorbent Assay, Incubation, Positive Control, Negative Control

    Binding activity of human dsNKG2D–IL-15 with MICA. ( a ) The human dsNKG2D–IL-15 protein was identified by the NKG2D antibody or IL-15 antibody through a western blot assay. ( b ) Plates coated with human dsNKG2D–IL-15, human NKG2D–Ig proteins, and IL-15 were bound by a human NKG2D antibody or an IL-15 antibody and were detected by ELISA. ( c ) Wells were coated with a soluble MICA protein (1 μg) or soluble RAE-1ε (1 μg). Human dsNKG2D–IL-15 was added and incubated at 37°C for 1 h. The NKG2D mAb, NKG2D pAb, or IL-15 pAb and HRP-conjugated secondary antibodies were added sequentially to the wells. The OD 450 value of each well was read after the addition of the substrate and stop solution. ( d ) All tumor cells were incubated with various concentrations of human dsNKG2D–IL-15, stained with a fluorescent IL-15 mAb and detected by flow cytometry. Gray area: isotype control; dotted line: IL-15 (5 μg mL −1 ) incubation and IL-15 mAb staining; and dashed line: human dsNKG2D–IL-15 incubation and IL-15 mAb staining. ( e ) B16-MICA cells were incubated with human or mouse dsNKG2D–IL-15 and detected with an IL-15 pAb, NKG2D mAb, or NKG2D pAb with or without MICA mAb neutralization. ( f ) B16BL6–MICA cells were co-incubated with serial concentrations of the human dsNKG2D–IL-15, mouse dsNKG2D–IL-15 or human NKG2D–Ig proteins (5 μg mL −1 ), stained with goat anti-human IgG antibody, and detected by flow cytometry. * P

    Journal: Cellular and Molecular Immunology

    Article Title: Human fused NKG2D–IL-15 protein controls xenografted human gastric cancer through the recruitment and activation of NK cells

    doi: 10.1038/cmi.2015.81

    Figure Lengend Snippet: Binding activity of human dsNKG2D–IL-15 with MICA. ( a ) The human dsNKG2D–IL-15 protein was identified by the NKG2D antibody or IL-15 antibody through a western blot assay. ( b ) Plates coated with human dsNKG2D–IL-15, human NKG2D–Ig proteins, and IL-15 were bound by a human NKG2D antibody or an IL-15 antibody and were detected by ELISA. ( c ) Wells were coated with a soluble MICA protein (1 μg) or soluble RAE-1ε (1 μg). Human dsNKG2D–IL-15 was added and incubated at 37°C for 1 h. The NKG2D mAb, NKG2D pAb, or IL-15 pAb and HRP-conjugated secondary antibodies were added sequentially to the wells. The OD 450 value of each well was read after the addition of the substrate and stop solution. ( d ) All tumor cells were incubated with various concentrations of human dsNKG2D–IL-15, stained with a fluorescent IL-15 mAb and detected by flow cytometry. Gray area: isotype control; dotted line: IL-15 (5 μg mL −1 ) incubation and IL-15 mAb staining; and dashed line: human dsNKG2D–IL-15 incubation and IL-15 mAb staining. ( e ) B16-MICA cells were incubated with human or mouse dsNKG2D–IL-15 and detected with an IL-15 pAb, NKG2D mAb, or NKG2D pAb with or without MICA mAb neutralization. ( f ) B16BL6–MICA cells were co-incubated with serial concentrations of the human dsNKG2D–IL-15, mouse dsNKG2D–IL-15 or human NKG2D–Ig proteins (5 μg mL −1 ), stained with goat anti-human IgG antibody, and detected by flow cytometry. * P

    Article Snippet: Then, the PVDF membranes were incubated with HRP-conjugated goat anti-rabbit IgG (1:3000 in TBST) (Invitrogen) for 1 h. The membranes were washed extensively with TBST, and immune reactive bands were visualized with a chemiluminescence reagent (Dakewe, Shenzhen, P.R.

    Techniques: Binding Assay, Activity Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation, Staining, Flow Cytometry, Cytometry, Neutralization

    Binding ELISA demonstrating the autoantibody binding to Met5A mesothelial cells due to the presence of MCAA in serum from human patients exposed to LA fibers. Sera were pooled from MCAA positive samples (MCAA+) and MCAA-positive samples cleared of all IgG (Cleared). Human mesothelial cells were plated in 96-well plates and then stained for MCAA binding using the pooled serum as the primary antibody and anti-human IgG - HRP for the secondary antibody. N= 3 samples per treatment group. Data are mean absorbance at 450 nm. *=p

    Journal: Inhalation toxicology

    Article Title: Mesothelial Cell Autoantibodies Upregulate Transcription Factors Associated with Fibrosis

    doi: 10.1080/08958378.2016.1271841

    Figure Lengend Snippet: Binding ELISA demonstrating the autoantibody binding to Met5A mesothelial cells due to the presence of MCAA in serum from human patients exposed to LA fibers. Sera were pooled from MCAA positive samples (MCAA+) and MCAA-positive samples cleared of all IgG (Cleared). Human mesothelial cells were plated in 96-well plates and then stained for MCAA binding using the pooled serum as the primary antibody and anti-human IgG - HRP for the secondary antibody. N= 3 samples per treatment group. Data are mean absorbance at 450 nm. *=p

    Article Snippet: The secondary antibody used was HRP-conjugated goat anti-rabbit IgG (ABCAM) diluted 1:1000 with 3%BSA/PBS.

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Staining