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ENZO ▼+FLUOR DE LYS® HDAC fluorometric activity assay kit

ENZO ▼+FLUOR DE LYS® HDAC fluorometric activity assay kit

브랜드 ENZO
품명 +FLUOR DE LYS® HDAC fluorometric activity assay kit
CAT. NO BML-AK500-0001 분류 Kit
규격 1 x 96wells 재고 -
소비자가(₩) 738,000 CAS. NO
온도 -80℃ 위험물
수입발주일 매주 목요일 오후 5시
주의사항 Store all components except the microtiter plate and instruction booklet at -70°C for the highest stability

Information

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. 

 

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
 
Contents: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








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