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. SIRT3 is a mitochondrial protein that plays a role in metabolic regulation and other cellular processes. It is the only human sirtuin with a direct genetic link with longevity. Additional research needs to be done to further elucidate the mechanism behind the involvement of SIRT3 in metabolic dysfunction, aging, cancer, and neurodegenerative disease. Cayman’s SIRT3 Direct Fluorescent Screening Assay Kit provides a convenient fluorescence-based method for screening SIRT3 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(e-acetyl)-AMC, is incubated with human recombinant SIRT3 along with its cosubstrate NAD⁺. Deacetylation sensitizes the substrate such that treatment with the developer in the second step releases a fluorescent product. The fluorophore can be analyzed with an excitation wavelength of 350-360 nm and an emission wavelength of 450-465 nm.

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