The assay kit is designed to measure the phosphotransferase activity of MAP (mitogen-activated protein) Kinase in immunoprecipitates and column fractions. Crude cell lysates may also be used but detergents/biochemicals contained in the cell lysis buffer may inhibit MAP Kinase activity. Furthermore, although three inhibitors are included with the kit, editors may suggest other unknown kinases found in crude lysates are responsible for MBP phosphorylation. The assay kit is based on phosphorylation of a specific substrate (myelin
basic protein, MBP). The phosphorylated substrate is then analyzed by immunoblot analysis, probing with a monoclonal Phospho-specific MBP antibody. The enzyme assay is rapid, convenient and specific for MAP Kinase.
Kit Components:
M2353A: Assay Dilution Buffer 2x1ml
M2353B: Mab Myelin Basic Protein, phosphorylated (MBP) 1x100ug
M2353C: Magnesium/ATP Cocktail 2x1ml
M2353D: MAP Kinase Substrate Cocktail II: 2x1ml
M2353E: MAP Kinase Inhibitor Cocktail: 1x1ml
Storage and Stability:
Store other components at -20C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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