A SIMPLE FLUOROMETRIC METHOD FOR CAMP - APPLICATION TO STUDIES OF BRAIN ADENYLATE-CYCLASE ACTIVITY

  • 1 January 1981
    • journal article
    • research article
    • Vol. 7  (1) , 27-35
Abstract
A simple fluorometric method for the determination of cAMP is presented. The fluorescent derivative is 1,N6-etheno adenosine 3'',5''-cyclic monophosphate (etheno-cAMP). Maximal formation of this derivative occurs after reacting cAMP with chloroacetaldehyde for 15 min at 100.degree. 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 [high pressure liquid chromatography] system. The derivatization as well as the fluorescent response of etheno-cAMP is linear between 2.5 and 700 pmol of cAMP. Studies of [guinea pig] brain adenylate cyclase by the fluorometric/HPLC method indicated that this method is comparable to the established radioenzymatic method. The present method provides a simple rapid nonradioactive means for the determination of adenylate cyclase activity.

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