cAMP Assays

descriptionPublicationkeyboard_double_arrow_right Article 01 Apr 2007 English Publisher:Cold Spring Harbor LaboratoryJournal:Cold Spring Harbor Protocols, volume 2,007, page pdb.prot4739 (issn: 1559-6095, eissn: 1559-6095,

Authors: Gouzel, Karimova; Daniel, Ladant;

doi: 10.1101/pdb.prot4739

pmid: 21357070

cAMP Assays

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Abstract

INTRODUCTIONThe cAMP assay provides a quantitative determination of the efficiency of the functional complementation between pairs of hybrid proteins. cAMP measurements are obtained using ELISA. 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 described here. 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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