Fluor-de-Lys^® HDAC fluorometric activity assay kit

First-to-market by the leader in Epigenetics research tools



BML-AK500-0001	1 Kit	428.00 USD

- Useful for assaying lysates, immunoprecipitates or inhibitor screening using the nuclear extract provided.
- Includes HeLa nuclear extract, a rich source of HDACs 1 & 2 for use as a positive control or as a source of HDAC activity for screening.
- Compatible with class I & IIb HDAC and sirtuins (with addition of NAD⁺).
- Includes enough reagent for 100-200 assays.

No radioactivity. No extractions. HTS friendly-mix and read on one 96-well plate. For class I and class II HDACs/sirtuins.
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Figure 1: Reaction Scheme of the HDAC Fluorescent Activity Assay. Deacetylation of the substrate sensitizes it to the developer, which then generates a fluorophore (symbol). The fluorophore is excited with 360 nm light and the emitted light (460 nm) is detected on a fluorometric plate reader.

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Product Specification

Alternative Name:	Histone deacetylase fluorescent assay kit

Quantity:	96 assays

Kit/Set Contains:	Nuclear Extract from HeLa Cells (human cerival cancer cell line) (Prod. No. BML-KI140) (100 µl; In 0.1M potassium chloride, 20mM HEPES/sodium hydroxide, pH 7.9, 20% (v/v) glycerol, 0.2mM ethylenediaminetetraacetic acid, 0.5mM dithiothreitol, 0.5Mm PMSF, prepared according to a modification of J.D. Dignam et al. (1983) and S.M. Abmayr et al. (1988)). Storage: -70°C, avoid freeze/thaw cycles Fluor de Lys^® Substrate (Prod. No. BML-KI104) (50 µl; 50mM in DMSO) Storage: -70°C Fluor de Lys^® Developer Concentrate (20x) (Prod. No. BML-KI105) (300 µl; 20x stock solution, dilute in assay buffer before use) Storage: -70°C Trichostatin A (HDAC Inhibitor) (Prod. No. BML-GR309-9090) (100 µl; 0.2mM in DMSO) Storage: -70°C Fluor de Lys^® Deacetylated Standard (Prod. No. BML-KI142) (30 µl; 10mM in DMSO) Storage: -70°C HDAC Assay Buffer (Prod. No. BML-KI143) (20 ml; 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chlroide) Storage: -70°C 1/2 volume microplate (Prod. No. BML-KI101) Storage: Room temperature 1/2 volume white microplate (Prod. No. BML-K571) Storage: Room temperature

Long Term Storage:	-80°C

Use/Stability:	Store all components except the microtiter plate and instruction booklet at -70°C for the highest stability. The HeLa Nuclear Extract, Prod. No. BML-KI140, must be handled with particular care in order to retain maximum enzymatic activity. Defrost it quickly in a RT water bath or by rubbing between fingers, then immediately store on an ice bath. The remaining unused extract should be refrozen quickly, by placing at -70°C. If possible, snap freeze in liquid nitrogen or a dry ice/ethanol bath. To minimize the number of freeze/thaw cycles, aliquot the extract into separate tubes and store at -70°C. The Fluor de Lys™ Substrate, Prod. No. BML-KI104, when diluted in Assay Buffer, may precipitate after freezing and thawing. It is best, therefore, to dilute only the amount needed to perform the assays of that day.

General Literature References

Histone deacetylase is a target of valproic acid-mediated cellular differentiation: N. Gurvich et al.; Cancer. Res. 64, 1079 (2004), Abstract;

Phosphorus-based SAHA analogues as histone deacetylase inhibitors: G.V. Kapustin et al.; Org. Lett. 5, 3053 (2003), Abstract;

Inhibition of silencing and accelerated aging by nicotinamide, a putative negative regulator of yeast sir2 and human SIRT1: K.J. Bitterman et al.; J. Biol. Chem. 277, 45099 (2002), Abstract;

Cloning and characterization of a histone deacetylase, HDAC9: X. Zhou et al.; PNAS 98, 10572 (2001), Abstract;

Coupling of histone deacetylation to NAD breakdown by the yeast silencing protein Sir2: Evidence for acetyl transfer from substrate to an NAD breakdown product: J.C. Tanny et al.; PNAS 98, 415 (2001), Abstract;

Identification of a class of small molecule inhibitors of the sirtuin family of NAD-dependent deacetylases by phenotypic screening: C.M. Groezigner et al.; J. Biol. Chem. 276, 38837 (2001), Abstract;

A phylogenetically conserved NAD+-dependent protein deacetylase activity in the Sir2 protein family: J.S. Smith et al.; PNAS 97, 6658 (2000), Abstract;

Acetylation and chromosomal functions: W.L. Cheung et al.; Curr. Opin. Cell Biol. 12, 326 (2000), Abstract;

Cloning and characterization of a novel human class I histone deacetylase that functions as a transcription repressor: E. Hu et al.; J. Biol. Chem. 275, 15254 (2000), Abstract;

Histone deacetylases: silencers for hire: H.H. Ng et al.; Trends Biochem. Sci. 25, 121 (2000), Abstract;

Isolation of a novel histone deacetylase reveals that class I and class II deacetylases promote SMRT-mediated repression: H.-Y. Kao et al.; Genes Dev. 14, 55 (2000), Abstract;

Role of NAD(+) in the deacetylase activity of the SIR2-like proteins: J. Landry et al.; Biochem. Biophys. Res. Commun. 278, 685 (2000), Abstract;

Silent information regulator 2 family of NAD- dependent histone/protein deacetylases generates a unique product, 1-O-acetyl-ADP-ribose: K.G. Tanner et al.; PNAS 97, 14178 (2000), Abstract;

The language of covalent histone modifications: B.D. Strahl et al.; Nature 403, 41 (2000), Abstract;

Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase: S. Imai et al.; Nature 403, 795 (2000), Abstract;

A new family of human histone deacetylases related to Saccharomyces cerevisiae HDA1p: W. Fischle et al.; J. Biol. Chem. 274, 11713 (1999), Abstract;

HDAC4, a human histone deacetylase related to yeast HDA1, is a transcriptional corepressor: P. A. Wade et al.; Mol. Cell Biol. 19, 7816 (1999), Abstract;

Identification of a new family of higher eukaryotic histone deacetylases. Coordinate expression of differentiation-dependent chromatin modifiers: A. Verdel et al.; J. Biol. Chem. 274, 2440 (1999), Abstract;

Three proteins define a class of human histone deacetylases related to yeast Hda1p: C.M. Groezigner et al.; PNAS 96, 4868 (1999), Abstract;

SAP30, a novel protein conserved between human and yeast, is a component of a histone deacetylase complex: Y. Zhang et al.; Mol. Cell 1, 1021 (1998), Abstract;

Targeted recruitment of the Sin3-Rpd3 histone deacetylase complex generates a highly localized domain of repressed chromatin in vivo: D. Kadosh et al.; Mol. Cell. Biol. 18, 5121 (1998), Abstract;

Transcriptional repression by UME6 involves deacetylation of lysine 5 of histone H4 by RPD3: S.E.C. Rundlett et al.; Nature 392, 831 (1998), Abstract;

Histone acetylation in chromatin structure and transcription: M. Grunstein; Nature 389, 349 (1997), Abstract;

Isolation and characterization of cDNAs corresponding to an additional member of the human histone deacetylase gene family: W. M. Yang et al.; J Biol Chem 272, 28001 (1997), Abstract;

A mammalian histone deacetylase related to the yeast transcriptional regulator Rpd3p: J. Taunton et al.; Science 272, 408 (1996), Abstract;

Transcriptional repression by YY1 is mediated by interaction with a mammalian homolog of the yeast global regulator RPD3: W.M. Yang et al.; PNAS 93, 12845 (1996), Abstract;

Enzymatic deacetylation of histone: A. Inoue et al.; Biochem. Biophys. Res. Commun. 36, 146 (1969), Abstract;