| Assay Method Information | |
| | Polθ ATPase Enzymatic Assay (0.5 nM Enzyme Concentration) |
| Description: | PolQ ATPase enzyme (1-894) at 0.5 nM is incubated with a 10-point concentration response of inhibitors for 15 min at rt in the following buffer: 50 mM Tris Cl pH 7.5, 10% glycerol, 5 mM DTT, 10 mM MgCl2, 0.1 mg/ml BSA. Following pre-incubation with inhibitors, DNA (Fork C) at 20 nM and ATP at 100 μM are added to start the reaction. Enzymatic reaction proceeds at rt for 180 min ATP consumption is measured using the ADP-Glo assay from Promega. Luminescence is read on the Envision and IC50 are determined using the variable slope 4-parameters equation.Oligos are annealed by heating at 9500 for 5 min in the following buffer (10 mM Tris-HCl pH7.5, 50 mM NaCl, 1 mM EDTA) and cooled to rt. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |