An assay for adenylate cyclase (EC 126.96.36.199) is described using adenylyl imidodiphosphate (AMP-PNP) as substrate. Methods are described for introducing the cyclase reaction mixture directly into a protein binding assay for the cyclic nucleotide without purification of cyclic AMP. Significant substrate depletion and regenerating systems are avoided by this method, and blank values are negligible. The assay is capable of reproducibly detecting adenylate cyclase activity with less than 5 μg of protein from rat cerebral cortex.
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