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MEK1 and MEK2, also called MAP kinase kinase and ERK activator kinase, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling both cell growth and differentiation. MEK1/2 is activated by a wide variety of growth factors and cytokines, and also by membrane depolarization and calcium influx, with the predominant activators being Raf kinases. MEK is involved in the MAPK signaling pathway of Ras-Raf-MEK-ERK. This signaling cascade is an important target in cancer therapy, making high-throughput study methods in demand.
Figure 1: Measurement of phosphorylated and total MEK1/2.
NIH/3T3 cells were cultured in 96-well plates and serum-starved for 16 hours. Cells were then treated with 20% FBS for 30 minutes and fixed. Total and phospho MEK1/2 were each assayed in triplicate using the phospho and total MEK1/2 antibodies included in the FACE MEK1/2 Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet).
Antibody Specificities
The phospho-MEK1/2 antibody is specific for phosphorylated MEK1/2 and was raised against a synthetic phospho-peptide corresponding to residues surrounding Ser217/221 of human MEK1/2. This antibody recognizes MEK1/2 only when phosphorylated at Ser217/221, and also reacts with MEK1/2 when singly phosphorylated at Ser217 but not at Ser221. It does not cross-react with related kinases. The total-MEK1/2 antibody recognizes MEK1/2 proteins regardless of the phosphorylation state./p>
