HDAC6 Fluorogenic Assay Kit

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Catalog #
50076
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Description

The Fluorogenic HDAC6 Assay Kit is a complete assay system designed to measure histone deacetylase 6 (HDAC6) activity for screening and profiling applications. In addition, the kit includes purified HDAC6 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC6 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.

 

HDAC Activity

Need us to run inhibitor screens or profile your compounds against HDAC6? Check out our Deacetylase/Sirtuin Screening Services.

This product has been cited 22 times.

Synonyms
histone deacetylase, HDAC6, 50076-1, 50076-2, HDAC-6
Product Info
Storage and Usage
Citations22
Assay Kit Format
Fluorogenic
Supplied As
Kit comes in a convenient 96-well or 384-well format, with all the reagents necessary for 100 or 384 fluorescent HDAC6 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

96 Reactions

Catalog # Component Amount Storage 
50006 HDAC6 human recombinant enzyme 9 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 in DMSO
(200 µM)
100 µl -20°C
50031 HDAC Assay Buffer 10 ml -20°C
79685 Black, low binding NUNC black microtiter plate 1 Room Temp

 

 













 

 

 

384 Reactions

Catalog # Component Amount Storage 
50006 HDAC6 human recombinant enzyme 9 µg -80°C

Avoid
freeze/
thaw
cycles!

50037 Fluorogenic HDAC substrate 3 (5 mM) 50 µl -80°C
50030 2x HDAC Developer (contains Trichostatin A)
(2 µM)
2 x 6 ml -80°C
50031 HDAC Assay Buffer 10 ml -20°C
79961 384-well black, low binding microtiter plate 1 Room Temp

 

UniProt #
Q9UBN7
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.