Anti-IRS1 (phospho Y896) 抗体 (ab4873)

Peptide Competition:  Extracts prepared from CHO-T cells transiently transfected with human IRS-1 and treated with 50 nM insulin for 5 minutes were resolved by SDSPAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4^oC, then were incubated with 0.50 µg/mL ab4873 antibody, following prior incubation with: no peptide (1), a generic phosphotyrosine containing peptide (2), the non-phosphopeptide corresponding to the immunogen (3), or the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4873 blocks the antibody signal, thereby demonstrating the specificity of the antibody.

Phosphatase Stripping: Extracts prepared from CHO-T cells transiently transfected with human IRS 1 and