FLUOR DE LYS® SIRT1 fluorometric drug discovery assay kit

Store all components except the microplates and instruction booklet at -80°C for the highest stability. The SIRT1 enzyme, (Prod. No. BML-SE239), 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 enzyme 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 into separate tubes and store at -80°C. The 5x Developer II (Prod. No. BML-KI176) can be prone to precipitation if thawed too slowly. It is best to thaw this reagent in a room temperature water bath and, once thawed, transfer immediately onto ice.

SIRT1 (Sirtuin 1, hSir2 SIRT1) (human, recombinant) (Prod. No. BML-SE239) (100 U; One U=1 pmol/min at 37°C, 250 μM, FLUOR DE LYS ® Substrate (Prod. No. BML-KI104), 500 μM NAD ; Recombinant enzyme dissolved in 25mM TRIS, pH 7.5, 100mM sodium chloride, 5mM dithiothreitol and 10% glycerol. See vial label for activity and protein concentrations Storage: -80°C; AVOID FREEZE/THAW CYCLES! FLUOR DE LYS ® SIRT1 , Deacetylase Substrate (Prod. No. BML-KI177) (100μl; 5mM solution in 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -80°C FLUOR DE LYS ® Developer II Concentrate (5x) (Prod. No. BML-KI176) (5 x 250 μL; 5x Stock Solution; Dilute in Assay Buffer before use Storage: -80°C NAD (Sirtuin Substrate) (Prod. No. BML-KI282) (500 μl; 50 mM β-Nicotinamide adenine dinucleotide (oxidized form) in 50mM TRIS/CL, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -80°C Nicotinamide (Sirtuin Inhibitor) (Prod. No. BML-KI283) (500μl; 50 mM Nicotinamide in 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride) Storage: -80°C Resveratrol (Sirtuin Activator) (Prod. No. BML-KI284) (10 mg; Solid MW: 228.2, soluble in DMSO or 100% ethanol (to 100mM) Storage: -80°C Suramin sodium (Sirtuin Inhibitor) (Prod. No. BML-KI285) (10 mg; Solid MW: 1429.2, soluble in water or assay buffer (to 25mM) ) Storage: -80°C FLUOR DE LYS ® Deacetylated Standard (Prod. No. BML-KI142) (30 μl; 10mM in DMSO) Storage: -80°C Sirtuin Assay Buffer (50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride, 1 mg/ml bovine serum albumin) (Prod. No. BML-KI286) (20 ml) Storage: -80°C 1/2 volume microplates (Prod. No. 80-2407) 1 clear and 1 white, 96-well Storage: Room temperature