HDAC1 Fluorogenic Assay Kit

Catalog #
50061
$465 *
Size: 96 reactions
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Description

The Fluorogenic HDAC1 Assay Kit is a complete assay system designed to measure histone deacetylase 1 (HDAC1) activity for screening and profiling applications. In addition, the kit includes purified HDAC1 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC1 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 HDAC1. 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 32 times.

Synonyms
histone deacteylase, HDAC1, HDAC-1
Product Info
Storage and Usage
Citations32
Assay Kit Format
Fluorogenic
Supplied As
It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC1 activity measurements.
Materials Required But Not Supplied

0.1% solution (1 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

Format
Catalog # Components Amount Storage 
50051 HDAC1 human recombinant enzyme 1 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
  Black, low binding NUNC black microtiter plate 1 Room Temp 

 

UniProt #
Q13547
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. Santo, L., et al., Blood. 2012 Mar 15;119(11):2579-89.
2. Bradner, J.E., et al., Nat Chem Biol. 2010 Mar;6(3): 238-243.