| Assay Method Information | |
| | In Vitro Evaluation |
| Description: | IC50 values were determined to evaluate the inhibitory activity of the test compound on human ATR kinase.ATR/ATRIP(h) was incubated in an assay buffer containing 50 nM GST-cMyc-p53 and Mg/ATP (concentration as required). The reaction was initiated by adding a Mg/ATP mixture. After incubation for 30 min at room temperature, a stop solution containing EDTA was added to terminate the reaction. Finally, the detecting buffer containing the d2-labeled anti-GST monoclonal antibody and the europium-labeled anti-phospho Ser15 antibody against phosphorylated p53 were added. Then, the plate in time-resolved fluorescence mode was read and homogeneous time resolution was performed.The fluorescence (HTRF) signal was determined according to the formula: HTRF=HTRF=10000×(Em665 nm/Em620 nm). |
| Affinity data for this assay | |
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