Anti-MEK3 + MEK6 (phospho S189) antibody (ab4759)

Western blot - MEK3 + MEK6 (phospho S189) 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 immunogen (2), a generic phosphoserine containing peptide (3), a generic phosphothreonine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 antirabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4759 blocks the antibody signal, thereby demonstrating the specificity of the antibody. Please note that the Mal-E-tagged fusion protein expressing MEK6 runs at 81 kDa.

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 4oC, 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 immunogen (2), a generic phosphoserine containing peptide (3), a generic phosphothreonine containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 antirabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4759 blocks the antibody signal, thereby demonstrating the specificity of the antibody. Please note that the Mal-E-tagged fusion protein expressing MEK6 runs at 81 kDa.