The Fluorogenic HDAC Assay Kit is a complete assay system designed to measure histone deacetylase (HDAC) class 1 activity in cell extracts, nuclear extracts, immunoprecipitates, and purified enzymes. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC 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 HDAC 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 a sample containing HDAC activity (e.g. HeLa nuclear extract or purified HDAC enzyme). The deacetylation sensitizes the substrate so subsequent treatment with the Lysine Developer produces a fluorophore that can then be measured using a fluorescence reader at 485 nm (excitation)/528 nm (emission).
Intended Use:Designed to measure histone deacetylase (HDAC) class 1 activity in cell extracts, nuclear extracts, immunoprecipitates, and purified enzymes.
Test Principle:The Fluorogenic HDAC 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.
Kit Components:HDAC2 recombinant human enzyme: 1x6ugFluorogenic HDAC substrate 2 (5mM): 1x50ul2x HDAC Developer (contains Trichostatin A) (50uM): 1x6mlTrichostatin A (200uM) 1x100ulHDAC assay buffer: 1x10mlBlack, low binding NUNC black microtiter plate: 1 plate
Storage and Stability:Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
