Figure 1. CDK3/CycE 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), 10 ng/50 µl PEG20,000, Substrate: Rb CTF, 5 µg /50 µl, recombinant CDK3/CycE: 2 Units/50 µl.
Figure 2. Dose dependence curve of CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, increasing amounts of CDK3/CycE 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 CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, 50 ng CDK3/CycE, 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 3. Peptide concentration dependence of CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, 50 ng of CDK3/CycE 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 1. Time course of CDK3/CycE kinase activity: DELFIA® data generated using Phospho-Rb (Ser807/811) Antibody #9308 to detect phosphorylation of substrate peptide (#1144) by CDK3/CycE kinase. In a 50 µl reaction, 50 ng CDK3/CycE and 1.5 µM substrate peptide were used per reaction. (DELFIA® is a registered trademark of PerkinElmer, Inc.)