HDAC1 Fluorogenic BioAssay™ Kit

The Fluorogenic HDAC1 BioAssay™ Kit is a complete assay system designed to measure histone deacetylase 1 (HDAC1) activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC1 activity measurements. In addition, the kit includes purified HDAC1 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC1 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 HDAC1. 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 from|acetyllysines in modified proteins. In the HDAC assay, fluorescent-dye molecules are attached to a peptide containing acetyllysine. Attachment to the peptide quenches the fluorescence of the dye. After treatment of the peptide with an HDAC, the reaction is mixed with a development solution that is specific for nonacetylated lysines. If the acetyl group has been removed from the lysine by the HDAC, this solution will release the dye allowing for fluorescence. Fluorescence is therefore directly related to HDAC activity.||Kit Components:|Low binding NUNC black microtiter plate, 1x1plate|HDAC1 human recombinant enzyme, 1x1ug|Fluorogenic HDAC substrate (5mM), 1x50ul|2x HDAC Developer (contains Trichostatin A) (50um), 1x6ml|Trichostatin A (200um) in DMSO, 1x100ul |HDAC 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.