Simple assay for high-throughput screening of SIRT1 inhibitors/activators
- Detection method- Fluorescence (Ex/Em = 400/505 nm)
- Application-Screening/characterizing/studying SIRT1 inhibitors/activators
SIRT1 or Sirtuin 1 is a member of the silent information regulator 2 family. SIRT1, an NAD-dependent histone deacetylase can deacetylate histones and a number of nonhistone substrates, including p53. It is predominantly localized in the nucleus of normal cells. In cancer cells however, it is predominantly localized in the cytoplasm. SIRT1 has been shown to regulate a wide range of cellular functions that affect metabolic homeostasis and aging. SIRT1 exerts anti-apoptotic, anti-oxidative, and anti-inflammatory effects against cellular injury, and protects the cells through the regulation of mitochondrial biogenesis, autophagy, and metabolism in response to the cellular energy and redox status. SIRT1 also promotes vasodilation and protects vascular tissues. Activation and inhibition of SIRT1 is being targeted for various diseases. Unlike other known protein deacetylases, which simply hydrolyze acetyl-lysine residues, the sirtuin-mediated deacetylation reaction hydrolyzes acetyl-lysine and NAD. This hydrolysis yields the deacetylated substrate, O-acetyl-ADP-ribose and nicotinamide, itself an inhibitor of sirtuin activity. In BioVision’s SIRT1 Inhibitor/Activator screening Kit, Sirtuin 1 deacetylates the substrate, followed by cleavage of the deacetylated substrate to release the fluorescent group, which is detected fluorometrically at Ex/Em = 400/505 nm. In the presence of SIRT1 inhibitor, deacetylation is impeded, preventing cleavage of the substrate and release of the fluorescent group. The SIRT1 activator enhances SIRT1 activity resulting in a higher fluorescent signal in comparison to the control. This kit provides a rapid, simple, sensitive, and reliable test, which is suitable for high-throughput screening of SIRT1 inhibitors/activators. Inhibitor control (Nicotinamide) is included to compare the efficacy of the test inhibitors.
