| Assay Method Information | |
| | BRG1 ATPase Inhibition Activity |
| Description: | All subsequent reagent additions were performed using a MultiFlo FX Multi-Mode Dispenser. Assay buffer was 20 mM HEPES pH 7.5, 1 mM MgCl2, 20 mM KCl, 1 mM DTT, 0.01% BSA, 0.005% Tween 20. 4 μL of 7.5 nM Brg1 ATPase-SnAC in assay buffer was added to the assay plate and incubated at RT for 5 min with compound. 2 μL of 195 μM ATP and 6 nM pCMV-dR8.91 plasmid in assay buffer was added to assay plate to initiate the reaction. The final concentrations of reagents were 5 nM Brg1 ATPase-SnAC, 65 μM ATP, and 2 nM pCMV-dR8.91 plasmid. The ATPase reaction was incubated at RT for 60 min. 3 μL of ADP-Glo reagent was added to stop the reaction and was incubated for 30 min at RT. 3 μL of Kinase detection reagent was added to the assay plate which was incubated for 90 min at RT. Plates were read with a 2103 Multilabel Envision reader using ultrasensitive luminescence detection. |
| Affinity data for this assay | |
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