SIRT1 FRET-Based Screening Ass

人类SIRT1是酵母Sir2的同源物，已被证明能调节p53肿瘤抑制因子的活性并抑制细胞凋亡。Cayman的基于SIRT1FRET的筛选检测法提供了一种方便的荧光法来筛选SIRT1抑制剂或激活剂。该程序仅需两个简单的步骤，均在同一微孔板中进行。在第一步中，将与荧光团和猝灭剂偶联的底物与人类重组SIRT1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导致猝灭剂和荧光团分离。使用335-345nm之间的激发波长和440-465nm之间的发射波长分析产生的荧光。Human SIRT1 is the homolog of yeast Sir2 and has been shown to regulate the activity of the p53 tumor suppressor and inhibit apoptosis. Cayman's SIRT1 FRET-based Screening Assay provides a convenient fluorescence-based method for screening SIRT1 inhibitors or activators. The procedure requires only two easy steps, both performed in the same microplate. In the first step, the substrate, which is coupled to the fluorophore and quencher, is incubated with human recombinant SIRT1 along with its cosubstrate NAD+. Deacetylation sensitizes the substrate such that treatment with the developer in the second step results in the separation of the quencher and fluorophore. The resulting fluorescence is analyzed using an excitation wavelength between 335-345 nm and emission wavelength between 440-465 nm.