| Assay Method Information | |
| | ATR Kinase Inhibition Assay |
| Description: | This experiment adopted Homogeneous time-resolved fluorescence technology (HTRF ) of Cisbio Company for assay.Experiment method: buffers were prepared as follows:Final Concentration of VolumeName Resource concentration stock solution (uL)HEPES (mM) Gibco, Cat # 15630-080 25 1000 75Brij35 Sigma, Cat # 9002-92-0 0.01% 1% 30BSA (mg/mL) Sigma, Cat # B2064-50G 1 100 30DTT (mM) Sigma, Cat # D0632-10G 5 1000 15Glycerol Sigma, Cat # G5516-500ML % 50% 60MnCl2 (mM) Sigma, Cat # 7773-01-5 10 1000 30H2O 2760Total (uL) 3000Preparation of solutions of the test compounds: The example compounds shown in the table below were prepared into solutions using DMSO as solvent, the stock solution typically being 10 mM. The maximum starting concentration in this experiment was 3 uM, DMSO 3-fold serial dilutions for concentrations, the serially diluted solution was added to each well of the corresponding 384-well plate (geriner bio-one, Cat #784075) to give final concentrations of 3000, 1000, 333, 111, 37, 12.3, 4.12, 1.37, 0.457, 0.152 nM, respectively, and the corresponding amount of DMSO was added to the additional wells to be used as negative or positive control wells, respectively. |
| Affinity data for this assay | |
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