Nordeen S K, Green P P, Fowlkes D M
DNA. 1987 Apr;6(2):173-8. doi: 10.1089/dna.1987.6.173.
We present a rapid, sensitive enzymatic assay for chloramphenicol acetyltransferase (CAT) that does not require chromatography, HPLC, or autoradiography. The assay is based on the use of an inexpensive substrate, tritiated acetate, instead of [14C]chloramphenicol. The method is adapted from one originally used by de Crombrugghe et al. (1973) and by Shaw (1975), but with simplifications appropriate for routine use. In our hands, the method is as sensitive as the customary thin-layer chromatography assay and is far more efficient for the performance of many assays, both in terms of labor and expense.
我们提出了一种用于氯霉素乙酰转移酶(CAT)的快速、灵敏的酶促测定法,该方法无需色谱法、高效液相色谱法或放射自显影。该测定法基于使用一种廉价的底物——氚标记的乙酸盐,而非[14C]氯霉素。该方法改编自最初由德克伦布鲁格等人(1973年)以及肖(1975年)使用的方法,但进行了适合常规使用的简化。在我们的操作中,该方法与传统的薄层色谱测定法一样灵敏,并且在人力和费用方面,对于进行许多测定而言效率要高得多。