| Assay Method Information | |
| | HAT Activity Inhibition |
| Description: | The HAT activity inhibitory ability of HAT inhibitors was evaluated using SensoLyte HAT(p300) Assay Kit (ANASPEC, AS-72172). Specifically, 7.5 μL of the compound of Examples 1 to diluted with assay buffer was added to 7.5 μL of recombinant p300 solution diluted 10 times with assay buffer, and the mixture was incubated at room temperature for 10 minutes. 7.5 μL of acetyl CoA solution diluted 10 times with assay buffer and 15 μL of histone H3 peptide diluted 10 times with assay buffer were added thereto, and the mixture was incubated at 37° C. for 30 minutes. 37.5 μL of Stop Solution was added to stop the reaction. 75 μL of p300 Developer solution diluted 50 times with assay buffer was added, and the mixture was incubated at room temperature for 30 minutes under light-shielded conditions. The fluorescence at 513 nm when irradiated with 389 nm excitation light was measured using a multiplate reader. Based on the measured fluorescence intensity, the IC50 value, which corresponds to the concentration of the compound that shows 50% inhibition of the enzyme reaction, was calculated. |
| Affinity data for this assay | |
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