Anti-MEK3 (phospho S189/T193) + MEK6 (phospho S207/T211) antibody (ab4759)

Peptide Competition: Extracts prepared from background extracts with Mal-E tagged fusion protein expressing MEK 6 left inactivated (6) or with MEKK added for activation (1-5) were resolved by SDS-PAGE on a 10% polyacrylamide 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 ab4759 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 6), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic hosphoserine-containing peptide (3), a generic phosphothreonine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab"e;)2 anti-rabbit IgG alkaline phosphatase and signals were detected. The data show that only the phosphopeptide corresponding to MEK3/6 [pSpT189/193]/[pSpT207/211] blocks the antibody signal, demonstrating the specificity of the antibody. Please note that the Mal-E-tagged fusion protein expressing MEK6 runs at ~81 kDa.