The Fluorogenic HDAC Class2a Assay Kit is a complete assay system designed to measure histone deacetylase (HDAC) class2a activity in cell extracts, nuclear extracts, immunoprecipitates, and purified enzymes. It comes in a convenient 96 format, with all the reagents necessary for 100 fluorescent HDAC activity measurements. In addition, the kit includes purified HDAC4 and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC Class 2a 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 class2a activity level. First, the HDAC fluorometric substrate is incubated with a sample containing HDAC activity (e.g. HeLa nuclear extract or purified HDAC4 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.
1. HDAC4 human recombinant enzyme: 1x0.1ug. Store at -70 degrees C.
2. Fluorogenic HDAC substrate (5mM): 1x50ul. Store at -70 degrees C.
3. 2x HDAC Developer (contains Trichostatin A) (50uM): 1x6ml. Store at -70 degrees C.
4. Trichostatin A (200uM): 1x100ul. Store at -20 degrees C.
5. Histone Deacetylase, Assay Buffer (HDAC): 1x10ml. Store at -20 degrees C.
6. Black, low binding NUNC black microtiter plate: 1 plate. Store at room temperature.
Suitable for studying enzyme kinetics and screening small molecular inhibitorsof HDAC class2a for drug discovery and HTS applications. Other applications not tested.
Storage and Stability:
Store buffer and standard at -20 degrees C. Store enzyme and substrate at -70 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.