Abstract
Adenosine 3′,5′-cyclic monophosphate (cAMP) is involved in a myriad of normal and pathological processes. Indeed, this cyclic nucleotide serves as a second messenger for the action of endogenous and exogenous agents in organisms ranging from bacteria to humans (1). The ubiquitous nature of cAMP has made its measurement essential to the study of numerous hormones, local mediators, neurotransmitters, drugs, and toxins. Methods for the estimation of cAMP include enzymatic radioisotopic displacement (2), high-pressure liquid chromatography (HPLC) (3), protein kinase activation (4), luciferin-luciferase bioluminescence (5), competitive protein binding (6), and immunoassay techniques (7– 10).
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1.
Radioimmunoassay (RIA), using a high specific activity adenosine 3′,5′-cyclic phosphoric acid 2′-0-succinyl-3-[125I]iodotyrosine methyl ester together with a second antibody that is bound to magnetizable polymer particles.
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2.
Enzyme immunoassay (EIA), involving the linking of succinyl cAMP to horseradish peroxidase (HRP) and combining this with stable second-antrbody-coated microtiter plates.
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3.
Homogeneous RIA (nonseparation), incorporating the novel method of scintillation proximity assay (SPA).
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References
Schram, M. and Selinger, Z. (1984) Message transmission\ receptor controlled adenylate cyclase system. Science 225, 1350–1356.
Butcher, R. W., Ho, R. J., Meng, H. C, and Sutherland, E. W. (1965) Adenosine 3′,5′-monophosphate in biological materials, The measurement of adenosine 3′,5′-mono-phosphate in tissues and the role of the cyclic nucleotides in the lipolytic response of fat to epinephrine. J. Biol. Chem. 240, 4515–4523.
Brooker, G. (1972) High-pressure anion exchange chromatography and enzymatic isotope displacement assays for cyclic AMP and cyclic GMP, in Advances in Cyclic Nucleotide Research, vol. 2 (Greengard, P, Paoletti, R., and Robinson, G. A., eds.), Raven, New York, pp. 111–129.
Kuo, J. K. and Greengard, P. (1970) Cyclic nucleotide-dependent protein kinases VIII. An assay method for the measurement of adenosine 3′,5′-monophosphate in various tissues and a study of agents influencing its level in adipose cells. J. Biol. Chem. 245, 4076–4083.
Ebadi, M. S., Weiss, B. and Costa, E. (1971) Microassay of adenosine 3′,5′-monophosphate (cyclic AMP) in brain and other tissues by luciferin-luciferase system. J. Neurochem. 18, 183–192.
Tovey, K. C, Oldham, K. G., and Whelan, J. A. M. (1974) A simple, direct assay for cyclic AMP in plasma and other biological samples using an improved competitive protein binding technique. Clin. Chim. Acta 56, 221–234.
Harper, J. F. and Brooker, G. (1975) Fentomole sensitive radioimmunoassay for cyclic AMP and cyclic GMP after 2′-0-acetylation by acetic anhydride in aqueous solution. J. Cyclic Nucleotide Res. 1, 207–218.
Steiner, A. L. (1974) Assay of cyclic nucleotides by radioimmunoassay methods, in Methods in Enzymology, vol. 38 (Langone, J J and Van Vunakis, H., eds.), Academic, London, pp. 96–105.
Cailla, H. L., Racine-Weisbuch, M. S., and Delaage, M. A. (1973) Adenosine 3′,5′cyclic monophosphate at 10-15 mole level. Anal. Biochem. 56, 394–407.
Tsugawa, M., Fida, S., Fujii, H., Moriwaki, K., Tarui, S., Sugi, M., Yamane, M., Yamane, R., and Fujimoto, M. (1990) An enzyme-linked immunosorbent assay (ELISA) for adenosine 3′,5′-cyclic monophosphate (cAMP) in human plasma and urine using monoclonal antibody. J. Immunoassay 11, 49–61.
Cailla, H. L., Roux, D., Kurtziger, H., and Delaage, M. A. (1980) Antibodies against cyclic AMP, cyclic GMP and cyclic CMP Their use in high performance radioimmunoassay. Hormones Cell Reg. 4, 1–25.
Steiner, A. L., Parker, S. W., and Kipnis, D. M. (1972) Radioimmunoassay for cyclic nucleotides; 1. Preparation of antibodies and iodinated cyclic nucleotides. J. Biol. Chem. 247, 1106–1113.
Scatchard, G. (1949) The attraction of proteins for small molecules and ions. Ann. NY Acad. Sei. 51, 660–672.
Erlanger, G. F. (1973) Principles and methods for the preparation of drug-protein conjugates for immunochemical studies Pharmacol. Rev. 25, 271–280.
Goldberg, N. D. and O’Toole, A. G. (1971) Analysis of cyclic 3′,5′-adenosine monophosphate and cyclic 3′,5′-guanosine monophosphate, in Methods of Biological Analysis, 20 (Glick, D, ed), Interscience Publishers, Wiley, London, pp. 1–39.
Steiner, A. L. (1979) Cyclic AMP and cyclic GMP, in Methods of Hormone Radioimmunoassay (Jaffe, B. M. and Behrman, H. R., eds.), Academic, New York, pp. 3–17.
Rosenberg, N., Pines, M., and Sela, I. (1982) Adenosine ′,5<limonophosphate—ts release in a higher plant by an exogenous stimulus as detected by radioimmunoassay. FEBS Lett. 137, 105–107.
Horton, J. K., Martin, R., Kalinka, S., Cushing, A., Kitcher, J. P., O’Sullivan, M. J., and Baxendale, P. M. (1992) Enzyme immunoassays for the estimation of adenosine 3′,5′-cychc monophosphate and guanosine 3′,5′-cyclic monophosphate in biological fluids J. Immunol. Methods 155, 31–40.
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© 1995 Humana Press Inc , Totowa, NJ
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Horton, J.K., Baxendale, P.M. (1995). Mass Measurements of Cyclic AMP Formation by Radioimmunoassay, Enzyme Immunoassay, and Scintillation Proximity Assay. In: Kendall, D.A., Hill, S.J. (eds) Signal Transduction Protocols. Methods in Molecular Biology™, vol 41. Humana Press. https://doi.org/10.1385/0-89603-298-1:91
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DOI: https://doi.org/10.1385/0-89603-298-1:91
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