Jenkins W T, Marshall M M
Anal Biochem. 1984 Aug 15;141(1):155-60. doi: 10.1016/0003-2697(84)90439-1.
A simple, rapid assay for purified ATPases is presented, based upon the formation of phosphomolybdate and its extraction into butyl acetate. The inclusion of imidazole makes the assay more sensitive and reproducible apparently because of the formation of an imidazole-phosphomolybdate complex. Protein (100 micrograms), Hepes buffer [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] (0.1 M) and nucleotides (1 mM) were all shown to cause interference. The interference by nucleotides could be counteracted by using more molybdate. Butyl acetate was shown to extract virtually all of the phosphomolybdate almost instantaneously upon vortex mixing.
本文介绍了一种基于磷钼酸盐的形成及其萃取到乙酸丁酯中的简单、快速的纯化ATP酶检测方法。加入咪唑后，该检测方法显然更灵敏且可重复，这可能是由于形成了咪唑 - 磷钼酸盐复合物。蛋白质（100微克）、Hepes缓冲液[4-(2-羟乙基)-1-哌嗪乙磺酸]（0.1M）和核苷酸（1mM）均显示会产生干扰。使用更多的钼酸盐可以抵消核苷酸的干扰。经涡旋混合后，乙酸丁酯几乎能立即萃取几乎所有的磷钼酸盐。
