Title: Quantitative monitoring of subcellular redox dynamics in living mammalian cells using roGFP2-based probes
Authors: Lismont, Celien
Walton, Paul A
Fransen, Marc # ×
Issue Date: 17-Apr-2017
Publisher: Humana Press, Inc.
Series Title: Methods in Molecular Biology vol:1595 pages:151-164
Abstract: To gain additional insight into how specific cell organelles may participate in redox signaling, it is essential to have access to tools and methodologies that are suitable to monitor spatio-temporal differences in the levels of different reactive oxygen species (ROS) and the oxidation state of specific redox couples. Over the years, the use of genetically-encoded fluorescent redox indicators with a ratiometric readout has constantly gained in popularity because they can easily be targeted to various subcellular compartments and monitored in real time in single cells. Here we provide step-by-step protocols and tips for the successful use of roGFP2, a redox-sensitive variant of the enhanced green fluorescent protein, to monitor changes in glutathione redox balance and hydrogen peroxide homeostasis in the cytosol, peroxisomes, and mitochondria of mammalian cells.
ISSN: 1064-3745
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Laboratory of Lipid Biochemistry and Protein Interactions
× corresponding author
# (joint) last author

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