HDAC3 Fluorogenic BioAssay™ Kit

The Fluorogenic HDAC3 BioAssay™ 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.

Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.

HDAC3 human recombinant enzyme, 1x2ug.
Fluorogenic HDAC substrate (5mM), 1x50ul
2x HDAC Developer (contains Trichostatin A) (50um), 1x6ml
Trichostatin A (200um), 1x100ul
HDAC assay buffer, 1x10ml
black, low binding NUNC black microtiter plate, 1x1plate

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. d

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.