Fluorogenic HDAC3 Assay Kit
Catalog #: 50073
DESCRIPTION: The Fluorogenic HDAC3 Assay Kit is a complete assay system
designed to measure histone deacetylase 3 (HDAC3) activity for screening and profiling
applications. It comes in a convenient 96-well format, with all the reagents necessary for
100 fluorescent HDAC3 activity measurements. In addition, the kit includes purified
HDAC3 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and
negative control. The Fluorogenic HDAC3 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 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.
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.
APPLICATIONS: Great for studying enzyme kinetics and screening small molecular
inhibitors for drug discovery and HTS applications.
STABILITY: One year from date of receipt when stored as directed.
1. Jia, H., et al., Neurobiology of Disease. 2012 May; 46(2): 351-361
2. Bradner, J.E., et al., Nat Chem Biol. 2010 Mar; 6(3): 238-243.