E.coli-derived human IRS1 recombinant protein (Position: S1041-Q1242). Human IRS1 shares 78% and 80% amino acid (aa) sequence identity with mouse and rat IRS1, respectively.
Conjugation:
Unconjugated
Alternative Names:
Insulin receptor substrate 1, IRS-1, IRS1
Anti-IRS1 Antibody (monoclonal, 10I3). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.
Clonality:
Monoclonal
Concentration:
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Application Notes: Western blot, 0.1-0.5µg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1µg/ml Immunocytochemistry/Immunofluorescence, 2µg/ml Flow Cytometry (Fixed), 1-3µg/1x106 cells. Add 0.2ml of distilled water will yield a concentration of 500µg/ml
Flow Cytometry analysis of PC-3 cells using anti-IRS1 antibody. Overlay histogram showing PC-3 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-IRS1 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-mouse IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Flow Cytometry analysis of U20S cells using anti-IRS1 antibody. Overlay histogram showing U20S cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-IRS1 Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-mouse IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
IF analysis of IRS1 using anti-IRS1 antibody. IRS1 was detected in immunocytochemical section of MCF7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-IRS1 Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of IRS1 using anti-IRS1 antibody. IRS1 was detected in paraffin-embedded section of human colon cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-IRS1 Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of IRS1 using anti-IRS1 antibody. IRS1 was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-IRS1 Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of IRS1 using anti-IRS1 antibody. IRS1 was detected in paraffin-embedded section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-IRS1 Antibody overnight at 4C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Western blot analysis of IRS1 using anti-IRS1 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug o
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