The Fluorogenic SIRT2 BioAssay™ Kit is a complete assay system designed to measure Sirtuin 2 (SIRT2) activity for screening and profiling applications. It comes in a convenient 96-well format, with all the reagents necessary for 32 fluorescent SIRT2 activity measurements. In addition, the kit includes purified SIRT2 enzyme and a SIRT inhibitor, Nicotinamide, for use as a positive and negative control, respectively. The kit includes sufficient HDAC substrate 1, buffers, NAD+, nicotinamide, and detection reagents for a full 96-well plate. Researchers can test their own SIRT2-containing samples in the remaining wells, or additional SIRT2 enzyme may be ordered separately. The Fluorogenic SIRT2 Assay Kit is based on a unique fluorogenic substrate and developer combination. This assay method eliminates dealing with the radioactivity, extraction, and chromatography aspects of traditional assays. Using this kit, only two simple steps on a microtiter plate are needed to analyze the SIRT2 activity level. First, the SIRT fluorometric substrate (SIRT substrate 1), containing an acetylated lysine side chain, is incubated with a sample containing purified SIRT2 enzyme. The deacetylation sensitizes the substrate so subsequent treatment with the Lysine Developer produces a fluorophore that can then be measured using a fluorescence reader.
Applications:Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Components:SIRT2 human recombinant enzyme, 1x135ugFluorogenic SIRT substrate 1, 1x50 ul (5mM)Nicotinamide Adenine Dinucleotide (NAD+) (50mM), 1x50ulNicotinamide (10mM), 1x500ul 2x SIRT Developer (contains 2mM Nicotinamide), 1x6ml HDAC assay buffer, 1x10mlblack, low binding NUNC black microtiter plate, 1 plate
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
