FLUOR DE LYS®-Green HDAC2 fluorometric drug discovery assay kit

• Useful for inhibitor screening or characterizing enzyme kinetics
• Includes optimal substrate selected from a panel of acetylated sites in p53 and histones
• Supplied with enough recombinant enzyme for 96 assays (1 x 96-well plate)

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.

HDAC2, like the highly homologous HDAC1, is a class I HDAC first identified as a human homolog of the yeast histone deacetylase Rpd3. HDAC2 found, along with HDAC1, in the Sin3, NuRD and CoREST, also can act independently of these complexes to deacetylate non-histone proteins such as transcription factors. In contrast to class II HDACs, which repress cardiac hypertrophy, HDAC2 activity is required for the cardiac hypertrophic response. This would suggest inhibition of HDAC2 as a possible therapeutic approach for heart disease. An opposite approach, i.e. increased HDAC2 activity or expression, might be indicated for chronic obstructive pulmonary disease (COPD), in which an HDAC2 deficit is implicated in insensitivity to glucocorticoids. The ability of the natural products theophylline and curcumin to increase HDAC2 expression is being investigated as an approach to this problem (see review). High HDAC2 expression is indicative of a poor prognosis in a number of cancers and, along with other HDACs, HDAC2 is a potential target for anti-cancer drug discovery.