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Research ArticleArticle

A SIMPLE, SENSITIVE METHOD FOR THE ASSAY OF ADENYL CYCLASE

GOPAL KRISHNA, BENJAMIN WEISS and BERNARD B. BRODIE
Journal of Pharmacology and Experimental Therapeutics October 1968, 163 (2) 379-385;
GOPAL KRISHNA
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BENJAMIN WEISS
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BERNARD B. BRODIE
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Abstract

A simple method for the amay of adenyl cyclase in tissue homogenates is described that uses either α-P82-, C14-or H3-labeled adenosine triphosphate (ATP) as substrate. Cyclic 3’, 5’- adenosine monophosphate (cyclic 3’, S’-AMP) formed during the incubation was isolated by chromatography on Dowex 50-H+ columns followed by precipitation of all nucleotides and inorganic phosphates by ZnSO4-Ba(OH)2, treatment, which left cyclic 3’, S’-AMP in the supernatant fluid. The purity of the cyclic 3’, 5’-AMP fraction was determined with the use of ion-exchange chromatography and paper chromatographic and electrophoretic techniques, as well as crystallization to constant specific activity. Furthermore, the purity of the nucleotide has been assessed enzymatically, by utilizing a purified cyclic nucleotide phosphodiesterase. The method described here makes possible the measurement of enzyme activities in tissue weighing less than 1 mg. Moreover, the entire procedure can be completed less than 3 hr.

Footnotes

    • Received March 25, 1968.
    • Accepted May 28, 1968.
  • © 1968 by the Williams & Wilkins Co.

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Journal of Pharmacology and Experimental Therapeutics
Vol. 163, Issue 2
1 Oct 1968
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Research ArticleArticle

A SIMPLE, SENSITIVE METHOD FOR THE ASSAY OF ADENYL CYCLASE

GOPAL KRISHNA, BENJAMIN WEISS and BERNARD B. BRODIE
Journal of Pharmacology and Experimental Therapeutics October 1, 1968, 163 (2) 379-385;

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Research ArticleArticle

A SIMPLE, SENSITIVE METHOD FOR THE ASSAY OF ADENYL CYCLASE

GOPAL KRISHNA, BENJAMIN WEISS and BERNARD B. BRODIE
Journal of Pharmacology and Experimental Therapeutics October 1, 1968, 163 (2) 379-385;
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