Green Sirt1 荧光检测试剂盒、Green SIRT1 Assay Kit *Fluorimetric*详细如下: Green SIRT1 Assay Kit 提供了一种方便的两步均相程序,用于使用荧光底物测量 Sirtuin 1 活性。在第一步中,乙酰化底物与含有 SIRT1 的样品一起孵育。底物的去乙酰化首先使其敏感,因此在第二步中,与 SIRT 显影剂混合会产生荧光团,然后可以在 Ex/Em= 490 nm/520 nm 处检测到。该试剂盒可用于 SIRT1 抑制剂/激活剂的高通量筛选。SIRT1 底物的长波长荧光受细胞成分和测试化合物的自发荧光的干扰较小。测定以标准的 96 孔格式进行。检测限:13 ng/ml 组蛋白去乙酰化酶 (HDAC) 酶通过组蛋白上赖氨酸残基的去乙酰化作为基因的转录抑制因子。Sirtuins (SIRT) 是一类独特的脱乙酰酶 (III 类 HDAC),需要 NAD 作为辅因子。 Add test compounds and SIRT1 enzyme to the microplate wells. For one assay in a 96-well plate, the suggested volume of enzyme solution is 40 µL and 10 µL of test compound. Establish the following control wells at the same time, as deemed necessary: Positive control contains SIRT1 enzyme without test compound. Inhibitor/activator control contains SIRT1 enzyme and SIRT1 inhibitor/activator. Vehicle control contains SIRT1 enzyme and vehicle used in delivering test compound (e.g. DMSO, concentration not to exceed 1%). Test compound control contains assay buffer (Component D) and test compound. Some test compounds may themselves be fluorescent and thereby give false results. Substrate control contains assay buffer (Component D). Using the assay buffer (Component D), bring the total volume of all controls to 50 µL. Pre-incubate the plate for 10 min at 37°C.