The Fluorogenic HDAC2 BioAssay™ Kit is a complete assay system designed to measure histone deacetylase 2 (HDAC2) activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC2 activity measurements. In addition, the kit includes purified HDAC2 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC2 Assay Kit is based on a unique fluorogenic substrate and developer combination. This assay method eliminates dealing with the radioactivity, extraction, and chromatography aspects of traditional assays. Using this kit, only two simple steps on a microtiter plate are needed to analyze the HDAC activity level. First, the HDAC fluorometric substrate, containing an acetylated lysine side chain, is incubated with purified HDAC2. The deacetylation sensitizes the substrate so subsequent treatment with the Lysine Developer produces a fluorophore that can then be measured using a fluorescence reader.
HDACs regulate cellular processes by catalyzing the hydrolysis of an acetyl group fromacetyllysines in modified proteins. In the HDAC assay, fluorescent-dye molecules areattached to a peptide containing acetyllysine. Attachment to the peptide quenches thefluorescence of the dye. After treatment of the peptide with an HDAC, the reaction ismixed with a development solution that is specific for nonacetylated lysines. If the acetylgroup has been removed from the lysine by the HDAC, this solution will release the dyeallowing for fluorescence. Fluorescence is therefore directly related to HDAC activity.
Applications:Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Kit Components:Low binding NUNC black microtiter plate, 1xplateHDAC2 human recombinant enzyme, 1x2ugFluorogenic HDAC substrate (5mM), 1x20ul2x HDAC Developer (contains Trichostatin A) (50uM), 1x6mlTrichostatin A (200um) in DMSO, 1x100ulHDAC Assay Buffer, 1x10ml
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
