SIRT1 Direct Fluorescent Scree

sirtuins代表一类独特的曲古抑菌素A不敏感的赖氨酰脱乙酰酶(III类HDAC)，可催化将赖氨酸脱乙酰化与烟酰胺和O-乙酰-ADP-核糖的形成偶联的反应。Cayman的直接荧光筛选检测试剂盒提供了一种方便的荧光筛选SIRT抑制剂或激活剂的方法。该过程仅需两个简单的步骤，均在同一微孔板中执行。在第一步中，将底物与人类重组SIRT及其共底物NAD+一起孵育。脱乙酰化使底物敏感，因此在第二步中用显影剂处理会释放荧光产物。可以使用350-360nm的激发波长和450-465nm的发射波长分析荧光团。The sirtuins represent a distinct class of trichostatin A-insensitive lysyl-deacetylases (class III HDACs) that catalyze a reaction coupling lysine deacetylation to the formation of nicotinamide and O-acetyl-ADP-ribose. Cayman's Direct Fluorescent Screening Assay Kits provide a convenient fluorescence-based method for screening SIRT inhibitors or activators. The procedure requires only two easy steps, both performed in the same microplate. In the first step, the substrate is incubated with human recombinant SIRT along with its cosubstrate NAD+. Deacetylation sensitizes the substrate such that treatment with the developer in the second step releases a fluorescent product. The fluorophore can be analyzed with an excitation wavelength of 350-360 nm and an emission wavelength of 450-465 nm.