| Assay Method Information | |
| | KAT6A Enzyme Activity Assay |
| Description: | a. The prepared enzyme solution was added to a 384-well plate at 3 μL/well, and 3 μL of 1× assay buffer was added to each well in columns 23 and 24 (Min).b. 3 μL of compound solution was added to each well, and 3 μL of buffer was added to each Min well; 3 μL of DMSO solution was added to each well in columns 1 and 2 (Max) as controls. The plate was centrifuged, shaken for 2 min, and incubated at room temperature for 15 min.c. The mixed substrate of Ac-CoA and H3 was added at 6 μL/well, and the plate was centrifuged, shaken for 2 min, and incubated at room temperature for 20 min.d. The assay reagent was added at 6 μL/well, and the plate was centrifuged, shaken for 2 min, and incubated in the dark at room temperature for 120 min.e. Plate reading was performed on the microplate reader, and AlphaScreen counts were recorded.f. Plotting was performed using the Graphpad software, and the IC50 values of the compounds were calculated. |
| Affinity data for this assay | |
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