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CS1010 Sigma-Aldrich

Histone Deacetylase Assay Kit, Fluorometric

sufficient for 100 assays (96 well plates)

Synonym: HDAC Assay Kit

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Properties

Related Categories Acetylation and Decacetylation Assays, Cell Biology, Cell Signaling and Neuroscience, Epigenetics, Gene Regulation and Expression,
Quality Level   200
usage   sufficient for 100 assays (96 well plates)
shipped in   dry ice
storage temp.   −20°C
Gene Information   human ... HDAC1(3065)

Description

Application

The Histone Deacetylase Assay Kit provides a simple method for the detection of HDAC activity based on a two-step enzymatic reaction. The substrate for this enzymatic assay is a substituted peptide with an acetylated lysine residue and a bound fluorescent group. The first step of the reaction is deacetylation of the acetylated lysine side chain by the HDAC containing sample (HeLa cell extract, purified enzyme, etc.). The second step is the cleavage of the deacetylated substrate by the developer solution and the release of the free highly fluorescent group. The measured fluorescence is directly proportional to the deacetylation activity of the sample.

Biochem/physiol Actions

Histone deacetylases (HDACs) are enzymes that remove acetyl groups from histone proteins. HDACs are important regulators of gene expression and genome function. They are implicated in a number of human disease states, namely several cancers, neurological disorders, and aging. Therefore, HDAC are relevant key targets for therapeutic intervention. Moreover, histone deacetylase inhibitors have been shown to serve as anti-tumor agents.

Features and Benefits

The kit includes all the reagents required for the fast and easy measurement of HDAC activity in cell or nuclear extracts, or with purified enzyme preparations. In addition, the kit provides HeLa cell lysate as a source of HDAC activity for inhibitor screening or as a positive control, a HDAC inhibitor (Trichostatin A), and a standard to enable activity quantitation.

Analysis Note

The Histone Deacetylase Assay Kit has been tested on HeLa, HEK 293T, NIH 3T3, and U 973 cell extracts.

Kit component only

Description

Product #

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Assay Buffer 20 mL    
Developer Solution 1.5 mL    
HDAC Inhibitor (Trichostatin A), 1 mM 50 μL    
HDAC Substrate, 20 mM 50 μL    
HeLa Cell Lysate 500 μL    
Standard (non-acetylated substrate), 4 mM 50 μL    
Safety & Documentation

Safety Information

RIDADR 
UN 1993 / PGIII
Flash Point(F) 
188.6 °F - closed cup
Flash Point(C) 
87 °C - closed cup
Protocols & Articles

Articles

Histone Modification and Chromatin Remodeling

Gene expression is governed by complex mechanisms including transcription factor binding to DNA and coordinated changes in chromatin structure. The primary protein components of chromatin are the his...
Savita Bagga, PhD.
BioFiles v7 n3, 2012, 10–16
Keywords: Acetylations, Amplification, Cancer, Cell culture, Chromatin immunoprecipitation, Cloning, DNA purification, Diseases, Epigenetics, Gene expression, Immunoprecipitation, Indicators, Methylations, Microarray Analysis, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymorphisms, Purification, Sequencing, Transcription, Whole genome amplification

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