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| | HDAC Activity Assay |
| Description: | A series of dilutions of the compounds were prepared with 10% DMSO in HDAC assay buffer, and 5uL of the dilution was added to a 50uL reaction so that the final concentration of DMSO is 1% in all of the reactions. The enzymatic reactions were conducted in duplicate at 37 degrees Celsius for 30 min in a 50uL mixture containing HDAC assay buffer, 5ug BSA, HDAC substrate, HDAC enzyme (human recombinant HDAC1, HDAC6, or HDAC8) and various concentrations of each compound. After enzymatic reactions, 50uL 2X HDAC Developer was added to each well and the plate was incubated at RT for an additional 15 min. Fluorescence intensity of measured at an excitation of 360nm and an emission of 460nm using a Biotek Synergy microplate reader. |
| Affinity data for this assay | |
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