SIRT-Glo™ Assay and Screening System Technical Manual

Literature # TM336

The SIRT-Glo™ Assay and Screening System are single-reagent-addition, homogeneous, luminescent assays that measure the relative activity of the NAD+-dependent histone deacetylase (HDAC) class III enzymes (sirtuins; SIRTs) from purified enzyme sources. This assay is broadly useful for class III enzymes, but sensitivity and performance will vary with catalytic efficiency of particular isoenzymes. The assay uses an acetylated, luminogenic peptide substrate that can be deacetylated by SIRT activities. Deacetylation of the peptide substrate is measured using a coupled enzymatic system in which a protease in the Developer Reagent cleaves the peptide from aminoluciferin, which is quantified in a reaction using Ultra-Glo™ Recombinant Luciferase. The SIRT-mediated luminescent signal is persistent and proportional to deactetylase activity, allowing batch processing of multiwell plates.