| Assay Method Information | |
| | CDK4 Activity Assay |
| Description: | CDK4 and cyclin D1 were expressed in a baculovirus expression system and subsequently purified. The catalytic activity of the CDK4 protein was assayed by measuring the phosphorylation of a truncated GST-Rb protein (residues 773-928) in the presence of ATP/ [gamma-33P] ATP. 33P incorporation was measured with a scintillation counter. IC50 values were determined by a least-squares fit to the equation, CPM = Vmax X(1 - ([I]/(IC50 + [I]))) + nonspecific binding. Ki,app = IC50/(1 + [ATP]/Km). |
| Affinity data for this assay | |
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