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

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



BML-AK500-0001	1 Kit	408.00 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

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;