Description:A fluorogenic, acetylated peptide substrate for SIRTs. It is a suitable substrate for SIRT1, SIRT2 and SIRT3 (tested so far). The substrate is based on residues 379-382 (Arg-His-Lys-Lys) of p53, a site of regulatory acetylation by the p300 and CBP acetyltransferases on lysines 381 and 382. This substrate fluoresces at excitation 350-380 nm, emission at 440-460 nm. The substrate should be used in conjunction with SIRT Developer. Supplied As: 5 mM DMSO solution (100 µl) Format: DMSO solution Instructions for use: Materials : SIRT Assay buffer (BPS catalog #50090) SIRT Assay developer (BPS catalog #50089) SIRT Substrate (BPS catalog #50080)SIRT1 (1 µg/µl) (BPS catalog #50012) or other SIRT enzyme Step 1 : Add the following reagents to a low binding NUNC black plate (VWR catalog #62408-936): SIRT Assay Buffer, 35 µl1 mg/ml BSA, 5 µl 200 µM SIRT substrate, 5 µlSIRT1, 5 µl Incubate at 37°C for 30 min. Step 2 : Stop the reaction by adding 50 µl of SIRT assay developer (undiluted) and incubate the plate at room temperature for 15 min. Step 3 : Read samples in a microtiter-plate reading fluorimeter capable of excitation at 350-380 nm and emission at 440-460 nm. \"Blank” value (no enzyme negative control) is subtracted from all other values. Storage / Stability:

Stable at least 12 months from date of receipt, when stored as directed

Application(s): Study enzyme kinetics, and screen small molecule sirtuin inhibitors for drug discovery and HTS applications. Reference(s):

1. A. Ito et al. (2001) EMBO J. 20 1331. 2. A. Ito et al. (2002) EMBO J. 21 6236.3. N.A. Barlev et al. (2001) Mol. Cell 8 1243.

Application Reference:SIRT2 Interacts with β-Catenin to Inhibit Wnt Signaling Output in Response to Radiation-Induced Stress (2014)

Warning(s): Protect from direct exposure to light. Avoid freeze/thaw cycles. Scientific Category: Deacetylase Product Type: Substrate Data shown is lot-specific. Contact us for specific information on other lots.