Use as a blocking reagent to evaluate the specificity of antibody reactivity in Western immunoblotting and dot blot protocols. For Western blotting, add 10 ul of antibody and 10 ul of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 30 minutes before allowing to react with the blot.
Supplied in 20 mM potassium phosphate (pH 7.0), 50 mM NaCl, 0.1 mM EDTA, 1 mg/ml BSA and 5% glycerol. 1% DMSO. Store at –20°C.
This peptide is used to specifically block Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb #2338 reactivity.
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb #2338 signal in both dot blot and Western blotting.
MEK1 and MEK2, also called MAPK or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation (1-3). Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221, located in the activation loop of subdomain VIII, by Raf-like molecules. MEK1/2 is activated by a wide variety of growth factors and cytokines and also by membrane depolarization and calcium influx (1-4). Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC-12 cells (4). MEK activates p44 and p42 MAP kinase by phosphorylating both threonine and tyrosine residues at sites located within the activation loop of kinase subdomain VIII.
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