HDAC Fluorogenic Assay Kit (GWB-4C8F93)


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Description: The HDAC Fluorescent Activity Assay kit is a complete assay system designed to measure histone deacetylase activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 uluorescent HDAC activity measurements. In addition, the kit includes purified HDAC2 and a potent HDAC inhibitor, TrichostatinA, for the positive and negative control. The HDAC Fluorescent Activity Assay is based on the 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 activity level. First, the HDAC fluorometric substrate, comprised of an acetylated lysine side chain, is incubated with a sample containing purified HDAC enzyme. The deacetylation will sensitize the substrate, causing further treatment with the Lysine Developer to produce a fluorophore that can then be measured using a fluorescence reader. COMPONENTS : Fluorogenic HDAC substrate: 50 ul @ 5 mM 2 x HDAC developer: 6 ml HDAC assay buffer: 10 ul Trichostatin A: 100 ul @ 200 uM One black plate 1 ug HDAC2 for the positive control APPLICATIONS : Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications. STORAGE : -20C or -70C. REFERENCES : 1. A. Ito et al., 2001, EMBO J. 20 1331. 2. N.A. Barlev et al., 2001, Mol. Cell 8 1243. 3. A. Ito et al., 2002, EMBO J. 21 6236. ASSAY PROTOCOL Material : Assay buffer, BPS catalog no. 50031, ; Assay developer, BPS catalog no. 50030, ; HDAC Substrate, catalog number 50037, Step 1 : Add all reaction mixture to a low binding NUNC black plate, VWR catalog no. 62408-936, 1 ul of HDAC2, 0.1 ug/ul, 39 ul of HDAC assay buffer, BPS catalog no. 50031, 5 ul of 1 mg/ml BSA 5 ul of 200 uM substrate, BPS catalog no. 50037, Incubate at 37 C for 30 min. Step 2 : Stop the reaction Add 50 ul of HDAC assay developer, 2x, BPS catalog no. 50030, and incubate the plate at room temperature for 15 min Step 3 : Read sample in a microtiter-plate reading fluorimeter capable of excitation at a wavelength in the range of 350-380 nm and detection of emitted light in the range of 440-460 nm.

Format: assay kit

Scientific Category: Deacetylase Assay

Additional Information

Name HDAC Fluorogenic Assay Kit (GWB-4C8F93)
Related Product Names histone deacetylase, HDAC
Purity NA
Format Assay Kit
Storage Mixed storage
Datasheets/Manuals Printable datasheet for GWB-4C8F93
Intended Use Research Use Only