背景介绍
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.
产品介绍
The Fluorogenic HDAC Assay Kit is a complete assay system
designed to measure histone deacetylase (HDAC) class 1 activity for screening and
profiling applications. 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 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.