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Analysis Note

The Histone Deacetylase Assay Kit has been tested on HeLa, HEK 293T, NIH 3T3, and U 973 cell extracts.

Application

The Histone Deacetylase Assay Kit provides a simple method for the detection of HDAC activity based on a two-step enzymatic reaction. The substrate for this enzymatic assay is a substituted peptide with an acetylated lysine residue and a bound fluorescent group. The first step of the reaction is deacetylation of the acetylated lysine side chain by the HDAC containing sample (HeLa cell extract, purified enzyme, etc.). The second step is the cleavage of the deacetylated substrate by the developer solution and the release of the free highly fluorescent group. The measured fluorescence is directly proportional to the deacetylation activity of the sample.

Biochem/physiol Actions

Histone deacetylases (HDACs) are enzymes that remove acetyl groups from histone proteins. HDACs are important regulators of gene expression and genome function. They are implicated in a number of human disease states, namely several cancers, neurological disorders, and aging. Therefore, HDAC are relevant key targets for therapeutic intervention. Moreover, histone deacetylase inhibitors have been shown to serve as anti-tumor agents.

Features and Benefits

The kit includes all the reagents required for the fast and easy measurement of HDAC activity in cell or nuclear extracts, or with purified enzyme preparations. In addition, the kit provides HeLa cell lysate as a source of HDAC activity for inhibitor screening or as a positive control, a HDAC inhibitor (Trichostatin A), and a standard to enable activity quantitation.