| Assay Method Information | |
| | Histone Deacetylase Assay (BPS Biosciences) |
| Description: | Histone deacetylase assay was done as per manufacturer's instructions. Briefly, assay buffer, 200 uM HDAC substrate (fluorogenic HDAC acetylated peptide substrate for class I HDACs (HDACs 1, 2 and 3) and class 2b HDACs (HDACs 6 and 10) and 1% BSA are taken as a master mixture and aliquoted as 40 uL per well. Compounds (10×) were diluted in assay buffer and were added to respective wells of a black 96 well plate. For HDAC1 (1.4 ng/μL) and for HDAC6 (7 ng/μL) human recombinant enzyme was thawed on ice and 5 μL of respective enzyme was added per well. The plate was incubated at 37° C. for 1 hour. Developer solution was then added (50 μL per well) and incubated at room temperature for 10 minutes. Fluorescence was measured at an excitation wavelength of 350-380 nm and emission wavelength of 440-480 nm. |
| Affinity data for this assay | |
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