一种使用反相高效液相色谱法的腺苷酸环化酶自动化检测方法。

An automated assay for adenylate cyclase using reversed-phase high-performance liquid chromatography.

作者信息

Reysz L J, Carroll A G, Jarrett H W

机构信息

Department of Biology, Purdue University School of Science, Indiana University, Indianapolis 46223.

出版信息

Anal Biochem. 1987 Oct;166(1):107-12. doi: 10.1016/0003-2697(87)90552-5.

DOI:10.1016/0003-2697(87)90552-5

PMID:3674402

Abstract

An automated high-performance liquid chromatographic method for the assay of 3',5'-cyclic AMP was developed using octylsilica. Total analysis time was 10.1 min, with cAMP eluting at 3 min. As little as 10 pmol of cyclic AMP could be detected by absorption at 260 nm. Peak height and area were linearly related to cyclic AMP concentration over at least two orders of magnitude. The analytical procedure gave good results in the assay of crude microsomal preparations of adenylate cyclase from both bovine brain and sea urchin eggs. The method was used to demonstrate that sea urchin adenylate cyclase is a Ca2+-activated enzyme.

摘要

开发了一种使用辛基硅胶的高效液相色谱自动化方法来测定3',5'-环磷酸腺苷（cAMP）。总分析时间为10.1分钟，cAMP在3分钟时洗脱。通过260nm处的吸光度可检测低至10皮摩尔的环磷酸腺苷。峰高和峰面积与环磷酸腺苷浓度在至少两个数量级范围内呈线性相关。该分析方法在测定牛脑和海胆卵腺苷酸环化酶的粗微粒体制剂时取得了良好结果。该方法用于证明海胆腺苷酸环化酶是一种Ca2+激活的酶。