SIRT6 or Sirtuin 6 proteins are a class of proteins that possess either histone deacetylase or mono-ribosyltransferase activity. SIRT6 is a nuclear sirtuin that has been associated with aging, cellular protection, sugar metabolism and certain types of cancer. Broad therapeutic applications are foreseen for SIRT6 inhibitors, including uses in diabetes, immune-mediated disorders, and cancer. Unlike other known protein deacetylases, which simply hydrolyze acetyl-lysine residues, the sirtuin-mediated deacetylation reaction hydrolyzes acetyl-lysine and NAD. This hydrolysis yields the deacetylated substrate, O-acetyl-ADP-ribose and nicotinamide, itself an inhibitor of sirtuin activity. Studies suggest that the human sirtuins may function as intracellular regulatory proteins with mono-ADP-ribosyltransferase activity. In Sirtuin 6 inhibitor screening Kit, Sirtuin 6 deacetylates the substrate, followed by cleavage of the deacetylated substrate to release the fluorescent group, which is detected fluorometrically at Ex/Em = 395/541 nm. In the presence of SIRT inhibitor, deacetylation is impeded, preventing cleavage of the substrate and release of the fluorescent group. This kit provides a rapid, simple, sensitive, and reliable test, which is suitable for high-throughput screening of SIRT6 inhibitors. Inhibitor control (Nicotinamide) is included to compare the efficacy of the test inhibitors.