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.
The Fluorogenic HDAC2 Assay 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.
histone deacteylase, HDAC2
DMSO >1%, strong acids or bases, ionic detergents, high salt
Instructions for use:
See assay kit data sheet for detailed protocol.
Storage / Stability:
12 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Great for studying enzyme kinetics and screening small molecular inhibitors of HDAC2 for drug discovery and HTS applications.
1. Santo, L., et al., Blood. 2012 Mar 15;119(11):2579-89.
2. Bradner, J.E., et al., Nat Chem Biol. 2010 Mar;6(3): 238-243.
MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:
0.1% solution (1 mg/ml) of bovine serum albumin (BSA) in water
Fluorimeter capable of excitation at 350-380 nm and detection at 440-460 nm
Adjustable micropipettor and sterile tips
Rotating or rocker platform