HDAC5 Fluorogenic Assay Kit

Catalog #

50065

$445 *

Size: 96 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The Fluorogenic HDAC5 Assay Kit is a complete assay system designed to measure histone deacetylase 5 (HDAC5) activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 100 fluorescent HDAC5 activity measurements. In addition, the kit includes purified HDAC5 enzyme and a potent HDAC inhibitor, Trichostatin A, for use as a positive and negative control. The Fluorogenic HDAC5 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 HDAC5 activity level. First, the HDAC fluorometric substrate, containing an acetylated lysine side chain, is incubated with purified HDAC5. The deacetylation sensitizes the substrate so subsequent treatment with the Lysine Developer produces a fluorophore that can then be measured using a fluorescence reader.

Need us to run inhibitor screens or profile your compounds against HDAC5? Check out our Deacetylase/Sirtuin Screening Services.

This product has been cited 9 times.

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Synonyms

histone deacteylase, HDAC5, HDAC-5

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Product Data Gallery

HDAC5 Activity

Product Info

Storage and Usage

Citations9

Assay Kit Format

Fluorogenic

Format

Catalog #	Component	Amount	Storage
50005	HDAC5 human recombinant enzyme	1 µg	-80°C	Avoid freeze/ thaw cycles
50040	Fluorogenic HDAC substrate class 2A (5 mM)	25 µl	-80°C
50030	2x HDAC Developer (contains Trichostatin A) (2 µM)	6 ml	-80°C
	Trichostatin A (1 mM) 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 #

Q9UQL6

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

Ontoria, J.M., et al., J. Med. Chem. 2009 Nov 12;52(21):6782-9.

Storage/Stability

6 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.

Instructions for Use

See assay kit data sheet for detailed protocol.

Applications

Useful for studying enzyme kinetics and screening small molecular inhibitors of HDAC5 for drug discovery and HTS applications.

Shipping Temperature

-80°C

Contraindications

DMSO >1%, strong acids or bases, ionic detergents, high salt

Warnings

Avoid freeze/thaw cycles.

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

Deacetylase

Regulatory Status

Research Use Only