Sinha A K, Colman R W
Biochem J. 1978 Sep 15;174(3):699-702. doi: 10.1042/bj1740699.
A method using the principle of affinity elution chromatography is described for the assay of adenylate cyclase in intact human platelets. By incubating platelet-rich plasma in the presence of radioactively labelled adenine, the ATP pool of the cells was prelabelled. Formation of labelled cyclic AMP from ATP was determined by extracting the platelets with HC1O4. After removal of the latter as KC1O4, the extract containing cyclic AMP and other adenine nucleotides was adsorbed in a NN-diethyl-N-2-hydroxypropylamino (QAE)-cellulose column. The column was washed, and subsequently cyclic AMP was specifically eluted with a cyclic AMP-dependent protein kinase and the radioactivity of the eluate was determined.
本文描述了一种利用亲和洗脱色谱原理来测定完整人血小板中腺苷酸环化酶的方法。通过在放射性标记的腺嘌呤存在下孵育富含血小板的血浆，细胞的ATP池被预先标记。用高氯酸提取血小板来测定由ATP形成的标记环磷酸腺苷。在以高氯酸钾形式除去高氯酸后，含有环磷酸腺苷和其他腺嘌呤核苷酸的提取物被吸附到NN-二乙基-N-2-羟丙基氨基（QAE）-纤维素柱上。对该柱进行洗涤，随后用一种环磷酸腺苷依赖性蛋白激酶特异性洗脱环磷酸腺苷，并测定洗脱液的放射性。
