HAT Inhibitor Screening Assay

Cayman的HAT抑制剂筛选试剂盒提供了一种快速的荧光法，用于评估PCAFHAT抑制剂。整个操作只需三个简单步骤，均在同一微孔板中完成。第一步中，HAT与乙酰辅酶A和组蛋白H3肽共同孵育，此时HAT催化乙酰基从乙酰辅酶A转移到H3肽上，生成乙酰化肽和辅酶A（CoASH）。在加入异丙醇终止酶反应后，将CPM加入孔板中，CPM与CoASH中的游离巯基反应，形成高荧光产物，其激发和发射波长分别为360-390nm和450-470nm。Cayman's HAT Inhibitor Screening Assay provides a fast, fluorescence-based method for evaluating PCAF HAT inhibitors. The procedure requires only three easy steps, all performed in the same microplate. In the first step of the protocol, HAT is incubated with acetyl-CoA and the histone H3 peptide. During this time, HAT catalyzes the enzymatic transfer of acetyl groups from acetyl-CoA to the H3 peptide producing an acetylated peptide and CoASH. Following addition of isopropanol to stop the enzymatic reaction, CPM is added to the wells of the plate. CPM reacts with the free thiol groups present on CoASH forming a highly fluorescent product that is detected using excitation and emission wavelengths of 360-390 and 450-470 nm, respectively.