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Histone deacetylase (HDAC) is an enzyme that removes acetyl group from a histone and plays an important role in regulating gene expression. Inhibition of HDAC can regulate transcription and induce apoptosis or differentiation in cancer cells. However, screening compounds that inhibit HDAC is difficult due to the lack of convenient way for detecting HDAC activity.
The HDAC Activity Fluorometric Assay Kit provides a fast and convenient way for detection of HDAC activity based on fluorescence method that eliminates radioactivity, chromatography or extractions in traditional assays. The assay needs only two easy steps performed on the same microtiter plate. First, the HDAC substrate that contains an acetylated lysine side chain is incubated with the sample containing HDAC activity (e.g., HeLa nuclear extract). Second, Deacetylation of the HDAC substrate sensitizes the substrate, so that further treatment with the Lysine Developer generates a fluorophore. The fluorophore can be easily detected using a fluorometer or a fluorescence plate reader. The assay is well suitable for high throughput screening applications.