| Assay Method Information | |
| | Enzyme Reaction System of CDK2/Cyclin A |
| Description: | The standard Lance Ultra method was performed by a 10 μL enzyme reaction system containing 0.5 nM CDK2/cyclin A protein, 100 nM ULight-MBP polypeptide, and 25 μM ATP. The dilutions of the compounds were dissolved in an enzyme buffer, respectively. The components of the buffer included 50 mM hydroxyethylpiperazine ethanesulfonic acid solution (pH 7.5), 1 mM ethylenediaminetetraacetic acid, 10 mM magnesium chloride, 0.01% Brij-35, and 2 mM dithiothreitol. After the reaction was started, an OptiPlate 384-well plate was sealed with a top heat-sealing film TopSeal-A and incubated at room temperature for 60 minutes. A stop buffer of the enzyme reaction was prepared, EDTA was dissolved in a 1-fold diluted detection buffer, and the reaction was terminated at room temperature for 5 minutes. 5 μL of the detection mixture (formulated with the europium-labeled anti-myelin basic protein antibody and the europium-labeled rabbit antibody, respectively) was added to the CDK2/cyclin A, CDK4/cyclin D1 and CDK6/cyclin D1 reaction systems, respectively. Incubation was carried out for 60 minutes at room temperature, and the reaction signals were detected using an Envision instrument according to the principle of time-resolved fluorescence resonance energy transfer. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |