The HDAC2 Fluorescent Activity Assay/Drug Discovery Kit is a complete assay system designed to measure the lysyl deacetylase activity of the recombinant human HDAC2 included in the kit. The kit is ideal for chemical library screening for candidate inhibitors or kinetic assay of the enzyme under varying conditions. The Fluor de Lys®-Green HDAC2 assay is based on the Fluor de Lys®-Green Substrate and Fluor de Lys® Developer combination. The assay procedure has two steps. First, the Fluor de Lys®-Green Substrate, which comprises an acetylated lysine side chain, is incubated with HDAC2. Deacetylation of the substrate sensitizes the substrate so that, in the second step, treatment with the Fluor de Lys®Developer produces a fluorophore. Fluor de Lys®-Green is an especially sensitive substrate for HDAC2. Activity can readily be measured with enzyme amounts in the range of 1-10 ng/well (0.36 – 3.6 nM in 50 ml), thus enabling IC50 determinations for high affinity inhibitors. The assay is highly suitable for high-throughput screening, with a Z’-factor of 0.74.
Study of enzyme kinetics, modulator activity, drug discovery.