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: |
05 Dec 2022 15:09 |
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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