A simple, sensitive method for the assay of adenyl cyclase.

摘要: 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’- monophosphate (cyclic S’-AMP) formed during incubation was isolated by chromatography on Dowex 50-H+ columns followed precipitation all nucleotides and inorganic phosphates ZnSO4-Ba(OH)2, treatment, which left cyclic S’-AMP supernatant fluid. The purity 5’-AMP fraction determined with use ion-exchange paper chromatographic electrophoretic techniques, well crystallization to constant specific activity. Furthermore, nucleotide has been assessed enzymatically, utilizing a purified phosphodiesterase. here makes possible measurement enzyme activities weighing less than 1 mg. Moreover, entire procedure can be completed 3 hr.