Extraction and Assay of Cyclic Nucleotides

  • Guy St. J. Whitley
Part of the Methods in Molecular Biology book series (MIMB, volume 27)

Abstract

Cyclic adenosine 3′,5′-monophosphate (cAMP) and cyclic guanosine 3′,5′-monophosphate (cGMP) have between them been implicated in the mediation of many physiological phenomena. Over the last 30 years, a number of techniques have been employed to assess changes in the amounts of these two molecules. Many, however, suffered from lack of sensitivity and/or ease of performance. Since the early 1970s, two methods have been developed that to a large extent overcome these problems. The first is based on competitive binding to endogenous proteins (1). The second method is a competitive radioimmunoassay that, with minor modifications, can be used to detect levels of cyclic nucleotides down to 1 fmol/assay tube; this is discussed in detail below (2,3). The method is equally applicable to the measurement of all cyclic nucleotides; however, for clarity only that for cAMP is described.

Keywords

Vortex Adenosine Polypropylene Anhydride Charcoal 

References

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Copyright information

© Humana Press Inc. Totowa, NJ 1994

Authors and Affiliations

  • Guy St. J. Whitley
    • 1
  1. 1.Department of Cellular and Molecular SciencesSt. George’s Hospital Medical SchoolLondonUK

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