The assay kit is designed to measure the phosphotransferase activity of Casein Kinase-2 (CK-2) in immunoprecipitates and column fractions. Crude cell lysates may also be used but detergents or biochemicals contained in the cell lysis buffer may inhibit CK-2 activity. Furthermore, although an inhibitor cocktail is included with the kit, editors may suggest other unknown kinases found in crude lysates are responsible for CK-2 substrate phosphorylation. The assay kit is based on phosphorylation of a specific substrate (CK-2 substrate peptide) using the transfer of the gamma-phosphate of [g-32P]ATP by CK-2 kinase. The phosphorylated substrate is then separated from the residual [g-32P]ATP using P81 phosphocellulose paper and quantitated by using a scintillation counter. The assay is linear for incubation times of up to 30 minutes and incorporation of up to 20% of total ATP. Further incubation or incorporation may not be linear and may therefore not be a true indication of CK-2 activity in the sample extract. The enzyme assay is rapid, convenient and fairly specific for CK-2. Each kit contains sufficient reagents for 200 individual CK-2 assays. detection and quantitation of casein kinase 2 activity from cell extracts.
Quantity:
~200 assays
Kit Components:
C2085-64A: Assay Dilution Buffer 2x1ml
C2085-64B: CK2 Substrate Peptide 2x1ml
C2085-64C: PKA Inhibitor Cocktail 2x1ml
C2085-64D: Mg/ATP Cocktail 2x1ml
C2085-64E: p81 pre-labeled squares 200
Storage and Stability:
Store at -20 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
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