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| | CK2 Phosphorylation Assay |
| Description: | Inhibitory effect of test compound was assessed in reaction buffer containing purified CK2, synthetic peptide substrate, and [gamma-33P]ATP. Assays were stopped by addition of 0.5 M orthophosphoric acid before spotting aliquots onto phosphocellulose filters. Filters were washed in 75 mM phosphoric acid (5-10 mL/each) four times then once in methanol and dried before counting. Inhibition constants were determined from Lineweaver-Burk double-reciprocal plots of data obtained in experiments run at fixed concentration of peptide substrate and at increasing concentrations of ATP either in the absence or in the presence of variable concentrations of inhibitor. Alternatively inhibition constants were also deduced from the IC50/Ki Cheng-Prusoff relationship. |
| Affinity data for this assay | |
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