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Cyclic AMP governs many fundamental signaling events in eukaryotic cells. Although cAMP signaling has been a major research focus for a long time, recent technological developments are revealing novel aspects of this paradigmatic pathway. In this chapter, we give an overview over current fluorescence resonance energy transfer (FRET)-based sensors for detection of cAMP dynamics, and their application in monitoring local, compartmentalized cAMP signals within living cells. A basic step-by-step protocol is given for conducting a FRET experiment in primary cells with a unimolecular cAMP sensor, which can easily be adapted to a user's specific requirements.

Original publication

DOI

10.1007/978-1-61779-126-0_16

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2011

Volume

746

Pages

297 - 316

Keywords

Automatic Data Processing, Biosensing Techniques, Cells, Cultured, Cyclic AMP, Fluorescence Resonance Energy Transfer, Intracellular Space, Myocytes, Cardiac, Signal Transduction