Adenylate cyclase assay with adenylyl imidodiphosphate and product detection by competitive protein binding

Michael E. Maguire, Alfred G. Gilman

Research output: Contribution to journalArticle

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Abstract

An assay for adenylate cyclase (EC 4.6.1.1) 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.

Original languageEnglish (US)
Pages (from-to)154-163
Number of pages10
JournalBBA - Enzymology
Volume358
Issue number1
DOIs
StatePublished - Jul 17 1974

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ASJC Scopus subject areas

  • Medicine(all)

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