Fluor-de-Lys™ HDAC fluorometric cellular activity assay kit

BML-AK503

Revised 01-Feb-10

Fluor-de-Lys™ HDAC fluorometric cellular activity assay kit
SYNONYMS	Cellular Histone deacetylase fluorescent assay kit
PRODUCT LINE	Epigenetics
PRODUCT CATEGORY	HDAC Kits & Sets

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
BML-AK503-0001		1 Kit	358.00 USD

Product Specification

QUANTITY:	96 assays
KIT/SET CONTAINS:	Nuclear Extracts from Hela Cells (human cervical cancer cell line) (Prod. No. BML-KI345) (50 µl; 2 mg protein/ml in 0.1M potassium chloride, 20mM HEPES/sodium hydroxide, pH 7.9, 20% (v/v) glycerol, 0.2mM ethylenediaminetetraacetic acid, 0.5M dithiothreitol, 0.1mM PMSF; Prepared according to a midification 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) (300 µl; 20x stock solution, dilutei n assay buffer before use) Storage: -70°C Trichostatin A (HDAC Inhibitor) (Prod. No. BML-GR-309-9090) (100 µl; 0.2mM in DMSO) Storage: -70°C Nicotinamide (Sirtuin Inhibitor) (Prod. No. BML-KI283) (500 µl; 50mM nicotinamide in 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -70°C Fluor de Lys™ Deacetylated Standard (Prod. No. BML-KI142) (30 µl; 10mM in DMSO) Storage: -70°C *HDAC Assay Buffer* (50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7 mM potassium chloride, 1mM magnesium chloride) (Prod. No. BML-KI143) (20 ml) Storage: -70°C Cell Lysis Buffer (Prod. No. BML-KI143) (20 ml; 1.0% NP-40 in HDAC Assay Buffer) Storage: -70°C 1/2 volume microplate (Prod. No. BML-KI101) Storage: Room temperature 1/2 volume white microplate (Prod. No. BML-KI110) Storage: Room temperature
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-80°C
USE/STABILITY:	Store all components except the microplate and instruction booklet at -70°C for the highest stability. The HeLa Nuclear Extract, Prod. No. BML-KI345, 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. BMLKI-104, 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.

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Product Description

The HDAC fluorometric cellular activity assay allows the determination of deacetylase activity within an undisturbed cellular environment and provides accurate activity information that is reflective of endogenous regulation. It also allows the study of the effects of upstream regulators on deacetylase activity and the detection of inhibitors or activators that act indirectly to affect deacetylase activity. The Fluor de Lys™ HDAC substrate is cell permeable. It is deacetylated by cellular HDACs and thus suitable for high-throughput cell-based deacetylase assays. This kit provides the reagents and protocols needed for determining rates of intracellular deacetylase activity with cultured cells. Deacetylation can be quantitated by addition of developer to the media and lysed cells.

General Information

HDACs are typically found in multiprotein complexes and are tightly regulated by subcellular localization, phosphorylation, and likely by other mechanisms.
BACKGROUND/TECHNICAL INFORMATION
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General Literature References

Acetylation: a regulatory modification to rival phosphorylation?: T. Kouzarides et al.; Embo J. 19, 1176 (2000) Abstract
Histone deacetylase inhibitors arrest polyglutamine-dependent neurodegeneration in Drosophila: J.S. Steffan et al.; Nature 413, 739 (2001) Abstract
Histone deacetylase inhibitors: P.A. Marks et al.; Adv. Cancer Res. 91, 137 (2004) Abstract
Suberoylanilide hydroxamic acid, a histone deacetylase inhibitor, ameliorates motor deficits in a mouse model of Huntington’s disease: E. Hockly et al.; PNAS 100, 2041 (2003) Abstract
Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan: K.T. Howitz; Nature 425, 191 (2003) Abstract
A novel action of histone deacetylase inhibitors in a protein aggresome disease model: L.J. Corcoran et al.; Curr. Biol. 14, 488 (2004) Abstract
Targeting CREB-bindinag protein (CBP) loss of function as a therapeutic strategy in neurological disorders: C. Rouaux et al.; Biochem. Pharmacol. 68, 1157 (2004) Abstract
Increased nuclear NAD biosynthesis and SIRT1 activation prevent axonal degeneration: T. Araki et al.; Science 305, 1010 (2004) Abstract
Sirt1 promotes fat mobilization in white adipocytes by repressing PPAR-gamma: F. Picard et al.; Nature 429, 771 (2004) Abstract
The novel histone deacetylase inhibitor BL1521 inhibits proliferation and induces apoptosis in neuroblastoma cells: A.J. de Ruijter; Biochem Pharmacol 68, 1279 (2004) Abstract
Sirtuin activators mimic caloric restriction and delay ageing in metazoans: J.G. Wood et al.; Nature 430, 686 (2004) Abstract
Modulation of NF-kappaB-dependent transcription and cell survival by the SIRT1 deacetylase: F. Yeung et al.; Embo J. 23, 2369 (2004) Abstract
Phase I study of an oral histone deacetylase inhibitor, suberoylanilide hydroxamic acid, in patients with advanced cancer: W. K. Kelly et al.; J. Clin. Oncol. 23, 3923 (2005) Abstract
Phase I and pharmacokinetic study of MS-275, a histone deacetylase inhibitor, in patients with advanced and refractory solid tumors or lymphoma: Q. C. Ryan et al.; J. Clin. Oncol. 23, 3912 (2005) Abstract
Resveratrol rescues mutant polyglutamine cytotoxicity in nematode and mammalian neurons: J.A. Parker et al.; Nat. Genet. 37, 349 (2005) Abstract
SIRT1 protects against microglia-dependent amyloid-beta toxicity through inhibiting NF-kappaB signaling: J. Chen et al.; J. Biol. Chem. 280, 40364 (2005) Abstract
Neuronal protection by sirtuins in Alzheimer's disease: T.S. Anekonda et al.; J Neurochem 96, 305 (2006) Abstract

Further Categories Containing This Product:

Activity Assays