Figure 1. CDK2/CycA kinase activity was measured in a radioisotopic filter binding assay using the following reaction conditions: 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, ATP (variable), 2.5 µg/50 µl PEG20,000, Substrate: Histone H1, 1 µg /50 µl, recombinant CDK2/CycA: 50 ng/50 µl.
Figure 3. Dose dependence curve of CDK2/CycA kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK2/CycA kinase. In a 50 µl reaction, increasing amounts of CDK2/CycA and 1.5 µM substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)
Figure 5. Staurosporine inhibition of CDK2/CycA kinase activity: DELFIA® data generated using Phospho-(Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK2/CycA kinase. In a 50 µl reaction, 50 ng CDK2/CycA, 1.5 µM substrate peptide, 20 µM ATP and increasing amounts of staurosporine were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)
Figure 4. Peptide concentration dependence of CDK2/CycA kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK2/CycA kinase. In a 50 µl reaction, 50 ng of CDK2/CycA and increasing concentrations of substrate peptide were used per reaction at room temperature for 30 minutes. (DELFIA® is a registered trademark of PerkinElmer, Inc.)
Figure 2. Time course of CDK2/CycA kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK2/CycA kinase. In a 50 µl reaction, 50 ng CDK2/CycA and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)