Anti-IRS1 (phospho S616) 抗体 (ab4776)

Extracts prepared from CHO-T cells mock transfected (Mock – lanes 1, 4), transiently transfected with wild-type human IRS 1 (WT – lanes 2, 5), and transiently transfected with mutant human IRS 1 (S616A – lanes 3, 6) and treated with TPA were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. After blocking non-specific sites with a 5% BSA-TBST buffer overnight at 4^oC, the membranes were incubated with 0.50 µg/mL ab4776 antibody (lanes 1-3), or an anti-IRS 1 pan antibody (lanes 4-6). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase. Signals were detected using the Tropix WesternStar method. The data show that the ab4776 antibody yields a signal on only the wild-type IRS 1 extract, not the mock or mutant IRS 1 extracts, thereby demonstrating the specificity of the antibody to this site. The data also show that the overall level of IRS 1 does not change between the wild-type and th