An assay for adenyl cyclase based on a combination of sodium borate electrophoresis and paper chromatography

Daniel Meruelo, Fred G. Bromberg, Michael A Edidin

Research output: Contribution to journalArticle

Abstract

We describe a method for the assay of adenyl cyclase in whole tissue homogenates. Adenosine 3′:5′-cyclic monophosphate (cAMP) formed from α-32P-, 14C- or 3H-labeled adenosine 5′-triphosphate (ATP) substrate is isolated from all known ATP metabolites and an unknown metabolite by electrophoresis in 1% sodium borate for 40 min, followed by overnight descending chromatography in 95% ethanol:1 m ammonium acetate (70:30). The purity of the cAMP isolated is established by chromatographic techniques as well as by utilizing a purified cyclic nucleotide phosphodiesterase. The method described here also makes possible the measurement of phosphodiesterase activity in homogenates. It is rapid enough to allow routine assay of 180 samples per day, although the number of samples processed depends on the number of electrophoretic and chromatographic units available.

Original languageEnglish (US)
Pages (from-to)30-43
Number of pages14
JournalAnalytical Biochemistry
Volume67
Issue number1
DOIs
StatePublished - 1975

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Paper Chromatography
Phosphoric Diester Hydrolases
Metabolites
Chromatography
Electrophoresis
Adenylyl Cyclases
Adenosine
Assays
Adenosine Triphosphate
Cyclic Nucleotides
Ethanol
Tissue
Substrates
sodium borate
triphosphoric acid
Cyclic AMP
ammonium acetate

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

An assay for adenyl cyclase based on a combination of sodium borate electrophoresis and paper chromatography. / Meruelo, Daniel; Bromberg, Fred G.; Edidin, Michael A.

In: Analytical Biochemistry, Vol. 67, No. 1, 1975, p. 30-43.

Research output: Contribution to journalArticle

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