The sirtuins represent a distinct class of trichostatin A-insensitive lysyl-deacetylases (class III HDACs) that catalyze a reaction coupling lysine deacetylation to the formation of nicotinamide and O-acetyl-ADP-ribose. The SIRT2 Direct Fluorescent Screening Assay provides a fluorescence-based method for screening SIRT2 inhibitors or activators. The procedure requires only two easy steps, both performed in the same microplate. In the first step, the substrate, which comprises the p53 sequence Gln-Pro-Lys-Lys(ε-acetyl)-AMC, is incubated with human recombinant SIRT2 along with its cosubstrate NAD+. Deacetylation sensitizes the substrate such that treatment with the Developer in the second step releases a fluorescent product.
SIRT2 Direct Assay Buffer (10X): 1 vial; -20°CSIRT2 (human recombinant): 2 vials; -80°CSIRT2 Direct Peptide: 2 vials; -20°CSIRT2 Direct NAD^+: 1 vial; -20°CSIRT2 Direct Nicotinamide: 1 vial; -20°C SIRT2 Direct Developer: 1 vial; -20°CSIRT2 Direct Fluorophore: 1 vial; -20°CHalf-Volume 96-Well Plate (white): 1 plate; Room temperature 96-Well Cover Sheet: 1 cover; Room temperature