Enzo Life Sciences Chemilum de Lys HDAC/SIRT Chemiluminescent Drug Discovery Kit

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The Chemilum de Lys HDAC/SIRT Chemiluminescent Drug Discovery Kit enables reproducible, high-throughput screening of HDAC or SIRT inhibitors using a novel substrate that closely mimics the natural in vivo substrate.

The 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 96-well format, with all reagents necessary for fluorescent HDAC activity measurements and calibration of the assay.

A HeLa nuclear extract, rich in HDAC activity, is included in 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 Chemilum de Lys Substrate and Developer combination. The Chemilum de Lys system (Chemiluminescent Histone deAcetylase Lysyl Substrate/Developer) is a sensitive and convenient alternative to radiolabelled, acetylated histones or peptide/HPLC methods for the assay of histone deacetylases.

The assay procedure has three steps. 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 sensitises the substrate so that, in the second step, treatment with the Chemilum de Lys Developer, followed by Enhancer, produces light.

Benefits include: luciferase-free reaction; high specificity; no interference from cell extract detergents; consistent results from a validated system; no artificial activation by resveratrol; resistance to protease inhibitors; resistance to kinase inhibitors.