Enzolution™ SIRT2 TR-FRET Assay System

The Enzolution™ SIRT2 Assay System is intended for use with the Transcreener® OAADPr SIRT Assay Kits (Parts #3046 and #3047) to measure the enzymatic activity of SIRT2 (NAD-dependent protein deacetylase sirtuin 2). SIRT2 catalyzes transfer of acetyl groups from lysine residues to NAD, yielding the unacetylated peptide, O-acetyl-ADP ribose (OAADPr), and nicotinamide as products (Figure 1). 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. 

The Enzolution SIRT2 Assay System provides all reagents required to screen and profile SIRT2 inhibitors when used with the Transcreener OAADPr SIRT Assay Kits, including purified, full-length human SIRT2 (aa 1-389) and acetylated peptide, 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.