SIRT1 Assay Kit

SIRT1 Assay Kit has been used in measuring the Sirt1 deacetylase activity in mesenchymal stem cells (MSCs), human retinal endothelial cells (HRECs) and human neuroblastoma SH-SY5Y cells.

Sirtuins (Sir2) are an evolutionarily conserved family of NAD⁺ dependent histone/protein deacetylases that tightly couple the cleavage of NAD⁺ and the deacetylation of protein substrates. The reaction products are nicotinamide, the deacetylated product, and a novel metabolite, 2′-O-acetyl-ADP-ribose. The proteins within this family are named after the first protein discovered from this family, Sir2 (Silent Information Regulator 2). Besides gene silencing, sirtuin proteins are important in other processes such as cell cycling regulation and fatty acid metabolism. SIRT1 is the human homolog of Sir2 and the one most studied to date. SIRT1 mediates p53 dependent process, transcription regulation, muscle differentiation, adipogenesis, and protection from axonal degeneration. SIRT1 also participates in early embryogenesis, neurogenesis, and cardiogenesis.

The kit offers all the reagents required for the fast and easy measurement of purified SIRT1 activity and for screening of inhibitors/activators. Moreover, the kit contains an inhibitor (nicotinamide) and an activator (resveratrol) as negative and positive controls, respectively.

The SIRT1 Assay Kit offers all the reagents required for the measurement of purified SIRT1 activity and for screening of inhibitors/activators.
The assay procedure is based on a two-step enzymatic reaction. The first step is deacetylation by SIRT1 of a substrate that contains an acetylated lysine side chain. The second step is the cleavage of the deacetylated substrate by the Developing Solution and the release of a highly fluorescent group. The measured fluorescence is directly proportional to the deacetylation activity of the enzyme in the sample.