| Assay Method Information | |
| | In Vitro Enzyme Inhibition Assay - HDAC |
| Description: | The effect of the compound on the enzymatic activity of pan-HDAC and two isoforms (HDAC1 and HDAC8) was examined using the HDAC-Glo I/II Assay and Screening System assay to assess the level of inhibition of HDAC protease activity and selectivity in each isoform. The proteins and detection reagents were from the HDAC-Glo™ I/II Assay and Screening System (Promega). A 1 mM stock solution of the compound was prepared in DMSO and diluted in a 5-fold gradient to create a series of 8 concentrations, and a specific volume of the highest concentration was diluted 25 times with HDAC Buffer to obtain 4× working solution of the compound. In a 384-well plate, 5 μL of the 4×HDAC enzyme solution and 5 μL of the test compound (4×) were sequentially added to each well, mixed evenly, and incubated at room temperature for 30 minutes. Subsequently 10 μL of 2× Developer reagent was added to each well, mixed evenly, and incubated at room temperature for 15-45 minutes. The 384-well plate was then read on a multifunctional microplate reader, with the Luminescence channel selected to record Luminescence values (RLU). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |