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Methods Mol Biol. 2015;1294:13-24. doi: 10.1007/978-1-4939-2537-7_2.

Recording intracellular cAMP levels with EPAC-based FRET sensors by fluorescence lifetime imaging.

Author information

1
Department of Cell Biology and Biochemistry, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands.

Abstract

Eukaryotic cells use second messengers such as Ca(2+), IP3, cGMP, and cAMP to transduce extracellular signals like hormones, via membrane receptors to downstream cellular effectors. FRET-based sensors are ideal to visualize and measure these rapid changes of second messenger concentrations in time and place. Here, we describe the use of EPAC-based FRET sensors to measure cAMP with spatiotemporal resolution in cells by fluorescence lifetime imaging (FLIM).

PMID:
25783874
DOI:
10.1007/978-1-4939-2537-7_2
[Indexed for MEDLINE]

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