Histone deacetylases (HDACs) are a large family of enzymes that remove acetyl groups from histone proteins. Site specific histone acetylation and deacetylation have been shown to activate or repress eukaryotic gene transcription, respectively, and as a consequence, HDACs play a crucial role in mammalian development and disease. HDACs are involved in important biological activities, such as cell differentiation, proliferation, apoptosis, and senescence. Of the several classes of HDACs, this kit assays a member of the zinc-independent, NADH requiring class, HDAC3. With Sigma′s HDAC3 Inhibitor Screening Kit, HDAC3 Enzyme acts with the supplied Developer to deacetylate and then cleave the HDAC3 Substrate [Arg-His-Lys-Lys(Ac)-AFC]. This activity releases the quenched fluorescent group, AFC, which can be detected at Em/Ex=380/500 nm. In the presence of a HDAC3 inhibitor, AFC is not released and its fluorescence remains quenched. The kit provides a rapid, simple, sensitive, and reliable test, suitable for either individual tests or high throughput screening of HDAC3 inhibitors. Trichostatin A (TSA) is included as a control inhibitor to compare with the efficacy of test inhibitors.
Features and Benefits
Simple, sensitive, and reliable assay
Simple procedure; takes ~60 min
Utilizes fluorometric methods
Sample type: candidate HDAC3 inhibitors
Suitable for screening HDAC3 inhibitors
Suitable for individual tests or high throughput assays and kinetic studies
Convenient 96-well microplate format
Other Notes
The kit is shipped on dry ice. Store HDAC-3 enzyme at -80 °C upon receipt. All other components should be stored at -20 °C, protected from light. All -20 °C reagents should be used within 2 months after thawing.
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