Detection and quantification of intracellular signaling using FRET-based biosensors and high content imaging

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

15 Citations (Scopus)

Abstract

Förster resonance energy transfer (FRET) biosensors represent invaluable tools to detect the spatiotemporal context of second messenger production and intracellular signaling that cannot be attained using traditional methods. Here, we describe a detailed protocol for the use of high content imaging in combination with FRET biosensors to assess second messenger production and intracellular signaling in a time-effective manner. We use four different FRET biosensors to measure cAMP levels, kinase (ERK and PKC), and GTPase activity. Importantly, we provide the protocols to express and measure these sensors in a variety of model cell lines and primary dorsal root ganglia neurons.
Original languageEnglish
Title of host publicationG Protein-Coupled Receptors in Drug Discovery
Subtitle of host publicationMethods and Protocols
EditorsMarta Filizola
Place of PublicationNew York NY USA
PublisherSpringer
Chapter10
Pages131-161
Number of pages31
Volume1335
Edition2
ISBN (Electronic)9781493929146
ISBN (Print)9781493929139
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Number2
Volume1335
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cell signaling
  • Förster resonance energy transfer
  • Biosensors
  • High content imaging
  • Extracellular-regulated kinase
  • Protein kinase C
  • Cyclic AMP
  • Rac1

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