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
Use/Stability
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.
