DESCRIPTION

• Notes: MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED: 1% solution (10 mg/ml) of bovine serum albumin (BSA) in water Fluorimeter capable of excitation at 350-380 nm and detection at 440-460 nm Adjustable micropipettor and sterile tips Rotating or rocker platform
• Uniprot: Q9BY41
• Shiptemp: -80°C (dry ice)
• Synonyms: histone deacetylase, HDAC
• Warnings: Avoid freeze/thaw cycles.
• Category: Deacetylase/Assay Kit
• Background: HDACs regulate cellular processes by catalyzing the hydrolysis of an acetyl group from acetyllysines in modified proteins. In the HDAC assay, fluorescent-dye molecules are attached to a peptide containing acetyllysine. Attachment to the peptide quenches the fluorescence of the dye. After treatment of the peptide with an HDAC, the reaction is mixed with a development solution that is specific for nonacetylated lysines. If the acetyl group has been removed from the lysine by the HDAC, this solution will release the dye allowing for fluorescence. Fluorescence is therefore directly related to HDAC activity.
• References: 1. Vijayakumar, B., et al. Bioinformation 2011; 7(3): 134-41. 2. Oehme, I., et al. Expert Opin Investig Drugs . 2009 Nov; 18(11): 1605-17. Application Reference(s): 1. Discovery of inhibitors of Schistosoma mansoni HDAC8 by combining homology modeling, virtual screening, and in vitro validation (2014) 2. Preclinical anti-arthritic study and pharmacokinetic properties of a potent histone deacetylase inhibitor MPT0G009 (2014)
• Description: The Fluorogenic HDAC 8 Assay Kit is a complete assay system designed to measure histone deacetylase (HDAC) 8 activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC activity measurements. In addition, the kit includes purified HDAC8 and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC 8 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 twoa simple steps on a microtiter plate are needed to analyze the HDAC 8 activity level. First, the HDAC fluorometric substrate is incubated with purified HDAC8 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.
• Methodology: Fluorogenic
• Unspsc Code: 41105331
• Unspsc Name: Protein chemifluorescent detection reagents or kits or substrates
• Applications: Great for studying enzyme kinetics and screening small molecular inhibitors of HDAC8 for drug discovery and HTS applications.
• Product Type: Assay Kit
• Biosafety Level: Not applicable (BSL-1)
• Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
• Storage Stability: 12 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
• Scientific Category: Deacetylase
• Instructions for Use: See assay kit datasheet for detailed protocol.