HDAC3 Fluorogenic Assay Kit

Catalog #
50073
$445 *
Size: 96 reactions
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Description

The Fluorogenic HDAC3 Assay Kit is a complete assay system designed to measure histone deacetylase 3 (HDAC3) activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC3 activity measurements. In addition, the kit includes purified HDAC3 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC3 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 activity level. First, the HDAC fluorometric substrate, containing an acetylated lysine side chain, is incubated with purified HDAC 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.

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This product has been cited 18 times.

Synonyms
histone deacetylase, HDAC3, HDAC-3
Product Info
Storage and Usage
Citations18
Assay Kit Format
Fluorogenic
Format
Catalog # Reagent Amount Storage
50003 HDAC3 human recombinant enzyme 2 µg -80°C Avoid freeze/ thaw cycles
50037 Fluorogenic HDAC substrate 3 (5 mM) 25 µl -80°C
50030 2x HDAC Developer (contains Trichostatin A) (2 µM)  6 ml -80°C
  Trichostatin A (200 µM) in DMSO  100 µl -20°C
50031 HDAC Assay Buffer 10 ml  -20°C
79685 black, low binding NUNC black microtiter plate 1 plate Room Temp

 

UniProt #
O15379
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. Jia, H., et al., Neurobiology of Disease. 2012 May; 46(2): 351-361
2. Bradner, J.E., et al., Nat Chem Biol. 2010 Mar; 6(3): 238-243.