Purified from rabbit serum by sequential epitope specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated MEK 7. The final product is generated by affinity chromatography using an MEK 7 derived peptide that is phosphorylated at serine 271 and threonine 275.
Dual specificity mitogen activated protein kinase kinase 7 antibody
Dual specificity mitogen-activated protein kinase kinase 7 antibody
JNK activating kinase 2 antibody
JNK kinase 2 antibody
JNK-activating kinase 2 antibody
JNKK 2 antibody
MAP kinase kinase 7 antibody
MAPK/ERK kinase 7 antibody
MAPKK 7 antibody
MEK 7 antibody
Mitogen Activated Protein Kinase kinase 7 antibody
MKK 7 antibody
PRKMK 7 antibody
SAPK kinase 4 antibody
Sek 2 antibody
stress-activated protein kinase kinase 4 antibody
Western blot - MEK7 (phospho S271 + T275) antibody (ab4762)
Predicted band size : 38 kDa
Peptide Competition: Extracts prepared from background extracts with tagged fusion protein expressing MEK 7 added 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 ab4762 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), 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 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method.
Phosphatase Stripping: Extracts were prepared as above. Membranes were left untreated (6, 8) or treated (7, 9) with lambda phosphatase, then incubated with 0.50 µg/mL ab4762 antibody (6, 7), or MEK 7 pan antibody (8, 9) for two hours at room temperature, and developed using the same procedure as above. The data show that only the peptide corresponding to ab4762 blocks the antibody signal, and that phosphatase stripping eliminates only the phospho- signal, thereby demonstrating the specificity of the antibody. Note that the tagged fusion protein expressing MEK 7 runs at 65 kDa.