Online Purchasing Account You are logged on as Guest. LoginRegister a New AccountShopping cart (Empty)
United States 

FLUOR DE LYS® HDAC fluorometric activity assay kit

First-to-market by the leader in Epigenetics research tools
 
BML-AK500-0001 96 wells 492.00 USD
Do you need bulk/larger quantities?
 
  • 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. Applications include cell-based assays and assay of immunoprecipitates.

Histone deacetylase inhibitors have shown promise as anti-tumor agents and naturally this has stimulated interest in the screening of compounds for HDAC inhibition. The FLUOR DE LYS® HDAC fluorometric activity assay kit is a sensitive and convenient alternative to protocols utilizing radiolabeled, acetylated histones or peptide/HPLC methods for the assay of histone deacetylases. It is based on the unique FLUOR DE LYS® (Fluorimetric Histone deAcetylaseLysyl) substrate and developer combination and provides an assay that can be carried out in two simple mixing steps, all on the same 96-well plate. First, the FLUOR DE LYS® substrate which comprises an acetylated lysine side chain, is incubated with a sample containing HDAC activity (HeLa nuclear or other extract, purified enzyme, bead bound immunocomplex, etc.). Deacetylation of the substrate sensitizes the substrate so that, in the second step, mixing with the FLUOR DE LYS® developer generates a fluorophore. The assay has been used successfully with preparations of all the known class I HDACs-HDAC1, HDAC2, HDAC3 and HDAC8 (see product data sheet) with class II HDACs 4-7, 9 and 10 and with the human Sir2 homolog, SIRT1 (see product data sheet). Work at Enzo Life Sciences has shown that the FLUOR DE LYS® substrate is cell-permeable and is deacetylated in situ by cellular HDACs. The deacetylated substrate accumulates inside cells and may be quantified by addition of FLUOR DE LYS® developer to a cell lysate.
bml-ak500

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.

Please mouse over
bml-ak500

Product Specification

Alternative Name:Histone deacetylase fluorescent assay kit
 
Applications:Fluorescent detection, HTS
Activity assay
 
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.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
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
 

Product Literature References

Adiponectin corrects premature cellular senescence and normalizes antimicrobial peptide levels in senescent keratinocytes: T. Jin, et al.; Biochem. Biophys. Res. Commun. 16, 31037 (2016), Application(s): Deacetylation activity measurement, Abstract;
Investigating the Sensitivity of NAD+-Dependent Sirtuin Deacylation Activities to NADH: A.S. Madsen, et al.; J. Biol. Chem. 291, 7128 (2016), Abstract; Full Text
Reversible Glutathionylation of Sir2 by Monothiol Glutaredoxins Grx3/4 Regulates Stress Resistance: N. Vall-Llaura, et al.; Free Radic. Biol. Med. 96, 45 (2016), Application(s): In vitro Sir2 activity was assayed, Abstract;
Hybrids from 4-anilinoquinazoline and hydroxamic acid as dual inhibitors of vascular endothelial growth factor receptor-2 and histone deacetylase: F.W. Peng, et al.; Bioorg. Med. Chem. Lett. 25, 5137 (2015), Application(s): Measurement of HDAC inhibitory activity , Abstract;
NBM-T-BBX-OS01, Semisynthesized from Osthole, Induced G1 Growth Arrest through HDAC6 Inhibition in Lung Cancer Cells: J.T. Pai, et al.; Molecules 20, 8000 (2015), Application(s): Assay using human H1299 cell lysates, Abstract; Full Text
Screening and profiling assays for HDACs and sirtuins: K.T. Howitz; Drug Discov. Today Technol. 18, 38 (2015), Abstract;
Molecular mechanism of sphingosine-1-phosphate action in Duchenne muscular dystrophy: D.H. Nguyen-Tran, et al.; Dis. Model Mech. 7, 41 (2014), Application(s): Assay, Abstract; Full Text
Synthesis and anticancer activities of thieno[3,2-d]pyrimidines as novel HDAC inhibitors: Q. Tan, et al.; Bioorg. Med. Chem. 22, 358 (2014), Application(s): Assay, Abstract;
Benzofused hydroxamic acids: useful fragments for the preparation of histone deacetylase inhibitors. Part 1: hit identification: E. Marastoni, et al.; Bioorg. Med. Chem. Lett. 23, 4091 (2013), Application(s): Assay, Abstract;
Design, synthesis and biological evaluation of di-substituted cinnamic hydroxamic acids bearing urea/thiourea unit as potent histone deacetylase inhibitors: C. Ning, et al.; Bioorg. Med. Chem. Lett. 23, 6432 (2013), Application(s): Assay, Abstract;
Dual-Acting Histone Deacetylase-Topoisomerase I Inhibitors: W. Guerrant, et al.; Bioorg. Med. Chem. Lett. 23, 3283 (2013), Application(s): Assay, Abstract; Full Text
A novel series of l-2-benzyloxycarbonylamino-8-(2-pyridyl)-disulfidyloctanoic acid derivatives as histone deacetylase inhibitors: design, synthesis and molecular modeling study: D. Huang, et al.; Eur. J. Med. Chem. 52, 111 (2012), Abstract;
Synthesis, evaluation and molecular modeling of cyclic tetrapeptide histone deacetylase inhibitors as anticancer agents: D. Huang, et al.; J. Pep. Sci. 18, 242 (2012), Abstract;
Activated microglia decrease histone acetylation and Nrf2-inducible anti-oxidant defence in astrocytes: restoring effects of inhibitors of HDACs, p38 MAPK and GSK3β: F. Correa, et al.; Neurobiol. Dis. 44, 142 (2011), Application(s): HDAC activity in astrocytes, Abstract;
Nuclear import of histone deacetylase 5 by requisite nuclear localization signal phosphorylation: T.M. Greco, et al.; Mol. Cell. Proteomics 10, M110.004317 (2011), Application(s): Activity of immunoprecipitated HDAC5, Abstract; Full Text
Rescue of the mutant CFTR chloride channel by pharmacological correctors and low temperature analyzed by gene expression profiling: E. Sondo, et al.; Am. J. Physiol. Cell. Physiol. 301, C872 (2011), Application(s): HDAC activity in lung cell lines, Abstract; Full Text
Drosophila SIN3 isoforms interact with distinct proteins and have unique biological functions: M.M. Spain, et al.; J. Biol. Chem. 285, 27457 (2010), Application(s): HDAC activity in nuclear extracts from S2 Drosophila cells, Abstract; Full Text
Sulforaphane Retards the Growth of Human PC-3 Xenografts and Inhibits HDAC Activity in Human Subjects: M.C. Myzak, et al.; Exp Biol Med 232, 227 (2007), Application(s): HDAC activity in PBMC lysates, Abstract; Full Text
Sulforaphane inhibits histone deacetylase in vivo and suppresses tumorigenesis in Apc-minus mice: M.C. Myzak, et al.; FASEB J. 20, 506 (2006), Application(s): HDAC activity in PBMC lysates, Abstract; Full Text
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;

General Literature References

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;
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.; BBRC 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;

Related Products

FLUOR DE LYS® HDAC fluorometric cellular activity assay kit 

Monitor HDAC activity in cell culture using cell-permeable FLUOR DE LYS® substrate
Fluorescent detection, HTS, Activity assay, Cell-based assays | Print as PDF
 
BML-AK503-0001 96 wells 445.00 USD
Do you need bulk/larger quantities?
 

FLUOR DE LYS®-Green HDAC fluorometric activity assay kit 

Fluorescent detection, HTS, Activity assay, Cell-based assays | Print as PDF
 
BML-AK530-0001 96 wells 404.00 USD
Do you need bulk/larger quantities?
 

FLUOR DE LYS® HDAC1 fluorometric drug discovery assay kit 

Fluorescent detection, HTS, Activity assay, Cell-based assays | Print as PDF
 
BML-AK511-0001 96 wells 656.00 USD
Do you need bulk/larger quantities?
 

FLUOR DE LYS® HDAC8 fluorometric drug discovery kit 

Fluorescent detection, HTS, Activity assay, Cell-based assays | Print as PDF
 
BML-AK518-0001 96 wells 656.00 USD
Do you need bulk/larger quantities?
 

FLUOR DE LYS® HDAC6 fluorometric drug discovery kit 

Fluorescent detection, HTS, Activity assay, Cell-based assays | Print as PDF
 
BML-AK516-0001 96 wells 656.00 USD
Do you need bulk/larger quantities?
 

FLUOR DE LYS®-Green HDAC2 fluorometric drug discovery assay kit 

Fluorescent detection, HTS, Activity assay, Cell-based assays | Print as PDF
 
BML-AK512-0001 96 wells 658.00 USD
Do you need bulk/larger quantities?
 

FLUOR DE LYS® HDAC3/NCOR1 fluorometric drug discovery kit 

Fluorescent detection, HTS, Activity assay, Cell-based assays | Print as PDF
 
BML-AK531-0001 96 wells 656.00 USD
Do you need bulk/larger quantities?
 

Product Toolbox

PRODUCT RESOURCES

Datasheet
MSDS
Certificate of Analysis
Manual

RELATED PRODUCTS

By target:
HDAC
By biological activity:
HDAC Activity kit
By catalog section:

PRODUCT SUPPORT

FAQs
Technical Service
Customer Service

Related Literature

Brochures
Stem Cells
Stem Cells
Download as PDF

Brochures
Epigenetics
Epigenetics
Download as PDF

Catalogs
Diabetes Catalog
Diabetes Catalog
Download as PDF

All new literature pieces

Recommend this page

 
For Research Use Only. Not for use in diagnostic procedures.
Keep in touch

©2017 Enzo Life Sciences, Inc.,