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Rapid assays for adenylate cyclase and 3',5' cyclic AMP phosphodiesterase activities. Simultaneous measurements of other pathways of ATP catabolism
Authors:M A Delaage  B N Bellon  H L Cailla
Institution:Centre de Biochimie et de Biologie Moléculaire-CNRS-31, chemin Joseph Aiguier-13274-Marseille-France
Abstract:This paper deals with a rapid assay of adenylate cyclase activity and 3′,5′ cyclic AMP phosphodiesterase activity which permits simultaneous measurements of other pathways of ATP catabolism. The separation of all the adenylic nucleotides was obtained by an electrophoresis on cellulose acetate 15 min at 50 V/cm with a fluorescent buffer pH 8.6. Before electrophoresis, the incubation sample was added with a carrier solution of nucleotides to allow their localization under uv light, by fluorescence inhibition. Each fraction was cut and dissolved in Bray's liquid for scintillation counting. This assay method is rapid, reliable, and sensitive, it is suitable either for research or for routine and clinical purposes.
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