一种简单的cAMP荧光测定方法:在脑腺苷酸环化酶活性研究中的应用。

W Wojcik, M Olianas, M Parenti, S Gentleman, N H Neff
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引用次数: 0

摘要

提出了一种简单的测定cAMP的荧光法。荧光衍生物为1,n6 -乙烯环3,5-单磷酸(乙烯- camp)。该衍生物的最大形成发生在cAMP与氯乙醛在100℃下反应15分钟后。荧光衍生物也从含有6-氨基嘌呤的化合物中产生。该方法的特异性在于使用反相/高效液相色谱系统。乙烯基cAMP的衍生化反应和荧光响应在cAMP的2.5 ~ 700皮摩尔之间呈线性关系。用荧光/高效液相色谱法对脑腺苷酸环化酶的研究表明,该方法与已建立的放射酶法相当。因此,本方法为测定腺苷酸环化酶活性提供了一种简单、快速、无放射性的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A simple fluorometric method for cAMP: application to studies of brain adenylate cyclase activity.

A simple fluorometric method for the determination of cAMP is presented. The fluorescent derivative is 1,N6-etheno cyclic 3,5-monophosphate (etheno-cAMP). Maximal formation of this derivative occurs after reacting cAMP with chloroacetaldehyde for 15 minutes at 100 degrees C. Fluorescent derivatives are also produced from compounds which contain a 6-amino purine. The specificity of the method resides in the use of a reverse phase/HPLC system. The derivatization as well as the fluorescent response of etheno-cAMP is linear between 2.5 and 700 picomoles of cAMP. Studies of brain adenylate cyclase by the fluorometric/HPLC method indicated that this method is comparable to the established radioenzymatic method. Thus, the present method provides a simple rapid nonradioactive means for the determination of adenylate cyclase activity.

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