| Contents | SIRT2 (Sirtuin 2, hSir2) (human, recombinant) (Prod. No. BML-SE25)
(500 U, one U=1 pmol/min at 37°C, 500µM, FLUOR DE LYS®-SIRT2 substrate (Prod. No. BML-KI179), 500µM 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®-SIRT2, Deacetylase substrate (Prod. No. BML-KI179)
(100µl; 5mM solution in 25mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassion chloride, 1mM magnesium chloride)
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; 5 mM Nicotinamide in 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride)
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 (Prod. No. BML-KI286)
(20 ml; 50mM TRIS/Cl, pH 8.0, 137mM sodium chloride, 2.7mM potassium chloride, 1mM magnesium chloride, 1 mg/ml bovine serum albumin)
Storage: -80°C
1/2 volume microplate (Prod. No. BML-KI101)
Storage: Room temperature
1/2 volume white microplate (Prod. No. BML-KI110)
Storage: Room temperature |
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| Use/Stability | Store all components except the microplates and instruction booklet at -80°C for the highest stability. The SIRT2 enzyme, Prod. No. BML-SE251, 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 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. |
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