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Selected Protocol from "Protein-Protein Interactions", 2nd Edition, E. Golemis, P. Adam.
Gouzel Karimova and Daniel Ladant
Unite Postulante de Biochimie des Interactions Macromoleculaires, Departement de Biologie Structurale et Chimie,
CNRS URA 2185, Institut Pasteur, Paris, France
Buy this book, Z719145
cAMP measurements are obtained using an ELISA assay (Harlow and Lane 1988). Commercial radio-immunoassays, or ELISA kits, to assay cAMP can be purchased from various manufacturers. In our laboratory we use the homemade, less-expensive ELISA that is described below. In outline, this assay is based on the ability of soluble cAMP in bacterial extracts to compete with binding of an alkaline phosphatase (AP)-conjugated anti-cAMP antibody to immobilized cAMP. The surface-bound, immobilized anti-cAMP-AP is inversely correlated to the concentration of cAMP in the extract. Known concentrations of soluble cAMP are used to calibrate the assay.
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