Enzolution™ SIRT1 TR-FRET Assay System

The Enzolution™ SIRT1 Assay System is intended for use with the Transcreener® OAADPr SIRT Assay Kits (Parts #3046 and #3047) to measure the enzymatic activity of SIRT1 (NAD-dependent protein deacetylase sirtuin 1). SIRT1 catalyzes transfer of acetyl groups from lysine residues to NAD, yielding the unacetylated peptide, O-acetyl-ADP ribose (OAADPr), and nicotinamide as products. The assay relies on a Coupling Enzyme (CE) to convert OAADPr into AMP, which is then detected using a far-red, competitive fluorescence polarization (FP) or time-resolved Förster-resonance-energytransfer (TR-FRET) assay. 

The assay has been optimized and extensively validated for high throughput screening (HTS) and inhibitor dose response measurements using most multimode plate readers.  Note that the assay was developed for endpoint detection; it has not been validated for use in continuous mode. The Enzolution SIRT1 Assay System provides all reagents required to screen and profile SIRT1 inhibitors when used with the Transcreener OAADPr SIRT Assay Kits, including purified, full-length human SIRT1 (aa 1-747) and acetylated peptide substrate, H3K9Ac (1-21). The protocol is configured for 384-well plates; use of different multi-well plate formats will require adjustment of reagent concentrations utilized in the assay. 

When purchasing the Enzolution Assay System, it is important to choose the assay system readout that corresponds to the TR-FRET configuration of your Transcreener OAADPr SIRT Assay.