| Assay Method Information | |
| | HAT Activity Inhibiting Assay |
| Description: | SensoLyte HAT (p300) Assay Kit (ANASPEC, AS-72172) was used to assess the HAT activity inhibiting capability of a HAT inhibitor. Specifically, 10 μL of compounds 1 to 6 and 14 to 19 diluted with an assay buffer was added to 10 μL of a recombinant p300 solution diluted 10-fold with an assay buffer, and the solution was incubated for 10 minutes at room temperature. 10 μL of acetyl-CoA solution diluted 10-fold with an assay buffer and 20 μL of histone H3 peptide diluted 10-fold with an assay buffer were added thereto, and the mixture was incubated for 30 minutes at 37° C. 50 μL of Stop Solution was added to quench the reaction. 100 μL of p300 Developer solution diluted 50-fold with an assay buffer was added, and the mixture was incubated for 30 minutes at room temperature under shaded conditions. 513 nm fluorescence when irradiated with 389 nm excitation light was measured using a multiplate reader. |
| Affinity data for this assay | |
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