| Assay Method Information | |
| | Inhibitory Activity Against CDK2 CycE1 Kinase |
| Description: | CDK2 CycE1 kinase solution (concentration: 0.015 ng/μL) was added to assay wells at 6 μL/well, and different compounds dissolved in DMSO were added to the assay wells separately using a nanoliter pipettor, such that the final concentrations of the compounds were 300 nM to 0.07 nM. The wells were set in duplicate, and a control well was set. After incubation of the above system for 30 min, ATP (concentration: 50 μM or 5000 μM) was mixed with ULight-Myelin Basic Protein Peptide substrate (manufacturer: PerkinElmer, concentration: 0.25 M) at a ratio of 1:1, and the mixture was added to the assay wells at 4 μL/well. The resulting mixture was left to react at room temperature for 2 h, and then 5 μL of EDTA was added to terminate the reaction, followed by addition of 5 μL of a detection antibody (manufacturer: PerkinElmer, concentration: 8 nM). The mixture was incubated at room temperature for 1 h. Assay was performed using a PerkinElmer Envision multifunctional microplate reader (excitation: 320 nm, emission: 615 nm/665 nm), and IC50 was calculated by four-parameter fitting. |
| Affinity data for this assay | |
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