The Fluorogenic HDAC4 Assay Kit is a complete assay system designed to measure histone deacetylase 4 (HDAC4) activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC4 activity measurements. In addition, the kit includes purified HDAC4 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC4 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 HDAC4 activity level. First, the HDAC fluorometric substrate, containing an acetylated lysine side chain, is incubated with purified HDAC4. 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.
References
Ontoria, J.M., et al., J. Med. Chem. 2009 Nov 12;52(21):6782-9.
Storage/Stability
6 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.
Instructions for Use
See assay kit data sheet for detailed protocol.
Applications
Useful for studying enzyme kinetics and screening small molecular inhibitors of HDAC4 for drug discovery and HTS applications.
Shipping Temperature
-80°C
Contraindications
DMSO >1%, strong acids or bases, ionic detergents, high salt
Warnings
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.