Detection of Phospho-MEK1 (T386) by Western Blot Western blot of human T47D cells showing specific immunolabeling of the ~45 kDa MEK1 (Control). The phosphospecificity of this labeling is shown in the second lane (lambda phosphatase, lambda PPase). The blot is identical to the control except that it was incubated in lambda PPase (1200 units for 30 minutes) before being exposed to the MEK1 (T386) antibody. The immunolabeling of MEK1 is completely eliminated by lambda PPase.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
For long-term storage, ≤ -20° C is recommended. Product is stable at ≤ -20° C for at least 1 year.
Human MEK1 (MAPK/ERK kinase 1) is a 45 kDa member of the MAP kinase kinase subfamily of the Ser/Thr protein kinase family of enzymes. It is 393 amino acids (aa) in length and contains one ERK interaction (aa 1 - 32) and catalytic site (aa 68 - 361). There is one alternate splice form that shows a 26 aa deletion between aa 471 - 548. MEK1 is part of the MAP kinase pathway that is activated in response to growth factors and integrins. Two signal transduction pathways converge on MEK1. Mitogens activate Ras, which activates Raf, which activates MEK1. Integrins activate Rac, which activates PAK, which also activates MEK1. Raf activates MEK1 by phosphorylating S218 and S222 on MEK1. Alternatively, PAK indirectly activates MEK1 by phosphorylating MEK1 on S298, which promotes the Raf phosphorylation of S218 and S222. Once activated, MEK1 phosphorylates and activates ERK1/2. In a negative feedback loop, activated ERK can phosphorylate MEK1 on T292 and T386. This phosphorylation of T292 will interfere with PAK phosphorylation of MEK1 on S298. ERK-mediated phosphorylation of MEK1 at T386 may also interfere with PAK phosphorylation of MEK1.
Chong, H. et al. (2003) Cell. Signal. 15:463.
Eblen, S.T. et al. (2004) Mol. Cell. Biol. 24:2308.
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