FLUOR DE LYS® HDAC fluorometric cellular activity assay kit

Store all components except the microplate at -80°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 -80°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 -80°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.

Nuclear Extracts from Hela Cells (human cervical cancer cell line) (Prod. No. BML-KI345) (50 μL; 2 mg protein/ml in 0.1M KCl, 20mM HEPES/NaOH, pH 7.9, 20% (v/v) glycerol, 0.2mM EDTAacid, 0.5M DTT, 0.1mM PMSF; Prepared according to a midification of J.D. Dignam et al (1983) and S.M. Abmayr et al. (1988) ) Storage: -80°C, avoid freeze/thaw cycles! FLUOR DE LYS ® Substrate (Prod. No. BML-KI104) (50 μL; 50mM in DMSO) Storage: -80°C FLUOR DE LYS ® Developer Concentrate (20x) (Prod. No. BML-KI105) (300 μl; 20x stock solution, dilute in assay buffer before use) Storage: -80°C Trichostatin A (HDAC Inhibitor) (Prod. No. BML-GR-309-9090) (100 μl; 0.2mM in DMSO) Storage: -80°C Nicotinamide (Sirtuin Inhibitor) (Prod. No. BML-KI283) (500 μl; 50mM nicotinamide in 50mM TRIS/Cl, pH 8.0, 137mM NaCl, 2.7mM KCl, 1mM MgCl2) Storage: -80°C FLUOR DE LYS ® Deacetylated Standard (Prod. No. BML-KI142) (30 μl; 10mM in DMSO) Storage: -80°C HDAC Assay Buffer (Prod. No. BML-KI143) (50mM TRIS/Cl, pH 8.0, 137mM NaCl, 2.7 mM KCl, 1mM MgCL2) (20 ml) Storage: -20°C Cell Lysis Buffer (Prod. No. BML-KI346 (20 ml; 1.0% NP-40 in HDAC Assay Buffer) Storage: -80°C 1/2 volume clear microplate (Prod. No. 80-2404) Storage: Ambient 1/2 volume clear microplate, sterile (Prod. No. 80-2430) Storage: Ambient