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Encompassing Amino Acids: Full Length
Application: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: CDK2/CyclinE2 activity was measured by using Histone H1 peptide (GGGPATP-KKAKKL-COOH), diluted in 20 mM Tris-HCl (pH 7.5) to a working concentration of 1 mg/ml, in the ADP Glo™ Kinase Assay kit (Promega #V9101). Reaction was initiated by mixing increasing amounts of CDK2/CyclinE2 with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl2, 0.1 mg/ml BSA prepared with 50 µM DTT, and substrate at a final concentration of 200 µg/ml.After a 40-minute incubation at room temperature, the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was added, and the reaction was incubated for another 30 minutes at ambient temperature. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate Multimode Reader. The Specific Activity was calculated as follows: (Corrected activity, RLU) / [(Specific activity from ADP in RLU/pmol) * (Reaction time in min) *(Enzyme amount in µg or mg)]. Corrected RLU was calculated by subtracting the blank value from all the values. The blank was determined from a "no enzyme" sample by replacing the enzyme solution with an equal volume of Kinase Dilution Buffer IX (1x).
Description: Complex of recombinant human full length CDK2 (cyclin dependent kinase 2) and CyclinE2. Both constructs contain an N-terminal GST-tag. The complex was affinity purified and is active.
Format: Aqueous buffer solution
Formulation: 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM Glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, and 25% glycerol
Genbank: CDK2: NM_001798
Cyclin E2: NM_057749
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tags
Uniprot: CDK2: P24941
Cyclin E2: O96020
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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