The HDAC/SIRT Chemiluminescent Drug Discovery Kit is a complete assay system designed to measure histone deacetylase (HDAC) and sirtuin activity in cell or nuclear extracts, immunoprecipitates or purified enzymes. It comes in a convenient 96-well format, with all reagents necessary for fluorescent HDAC activity measurements and calibration of the assay. In addition, a HeLa nuclear extract, rich in HDAC activity, is included with the kit. The extract is useful as either a positive control or as the source of HDAC activity for inhibitor/drug screening. Also included are Trichostatin A and Nicotinamide, which may be used as model inhibitors for HDACs and sirtuins, respectively.
The HDAC Chemiluminescent Activity Assay is based on the unique Chemilum de Lys™ Substrate and Developer combination.
The Chemilum de Lys™ system (Chemiluminescent Histone deAcetylase Lysyl Substrate/Developer) is a highly sensitive and convenient alternative to radiolabeled, acetylated histones or peptide/HPLC methods for the assay of histone deacetylases. The assay procedure has three steps (Fig. 1). First, the Chemilum de Lys™ Substrate, which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (HeLa nuclear or other extract, purified enzyme, bead-bound immunocomplex, etc.). Deacetylation of the substrate sensitizes the substrate so that, in the second step, treatment with the Chemilum de Lys™ Developer followed by Enhancer produces light. The reaction is luciferase-free.
Manufacturer Enzo Life Sciences
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