Measuring ERK Activity Dynamics in Single Living Cells Using FRET Biosensors.

Blum, Yannick; Fritz, Rafael D; Ryu, Hyunryul; Pertz, Olivier (2017). Measuring ERK Activity Dynamics in Single Living Cells Using FRET Biosensors. In: Jimenez, Gerardo (ed.) ERK signaling. Methods in molecular biology: Vol. 1487 (pp. 203-221). Springer 10.1007/978-1-4939-6424-6_15

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Fluorescence resonance energy transfer (FRET)-based biosensors are powerful tools for measuring spatio-temporal signaling dynamics in single living cells with subcellular resolution. There are quite a number of already existing sensors and this technology is increasingly used to obtain quantitative dynamic datasets. In this chapter, we describe the analysis of endogenous extracellular signal-regulated kinase (ERK) activity in living cells using the EKAR2G (ERK activity reporter second generation) probe. We focus on the generation of stable cell lines expressing the EKAR2G sensor as well as data acquisition and analysis.

Item Type:

Book Section (Book Chapter)

Division/Institute:

08 Faculty of Science > Department of Biology > Institute of Cell Biology

UniBE Contributor:

Pertz, Olivier

Subjects:

500 Science > 570 Life sciences; biology

ISSN:

1940-6029

Series:

Methods in molecular biology

Publisher:

Springer

Language:

English

Submitter:

Olivier Pertz

Date Deposited:

19 Feb 2018 15:48

Last Modified:

20 Mar 2019 07:53

Publisher DOI:

10.1007/978-1-4939-6424-6_15

PubMed ID:

27924569

Uncontrolled Keywords:

Biosensors Cell fate Cell-cell variability EKAR2G ERK dynamics Fluorescence resonance energy transfer (FRET) High throughput Live-cell microscopy PC12 cells Ratiometric FRET imaging Signaling heterogeneity

BORIS DOI:

10.7892/boris.109212

URI:

https://boris.unibe.ch/id/eprint/109212

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