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NADPH Oxidases pp 275-282 | Cite as

Imaging Intracellular H2O2 with the Genetically Encoded PerFRET and OxyFRET Probes

  • Balázs EnyediEmail author
  • Miklós GeisztEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1982)

Abstract

Hydrogen peroxide (H2O2) is an important signaling intermediate with various regulatory and effector functions. Despite its significance, the subcellular organization of H2O2 signals is poorly understood. Introducing novel techniques for the intracellular detection of H2O2 would be essential for a more complete understanding of its role in cellular signaling. We previously reported the development of two novel fluorescence resonance energy transfer (FRET)-based protein sensors that showed opposite emission ratio changes upon reaction with H2O2. In this chapter, we detail the methods for using OxyFRET and PerFRET for the assessment of changes in subcellular H2O2 levels.

Key words

Hydrogen peroxide PerFRET OxyFRET Genetically encoded sensor Fluorescent biosensor imaging 

Notes

Acknowledgment

This work was supported by grants from the Hungarian Research Fund (OTKA NF72669) and from the National Office for Research and Technology (OMFB-01680/2009). The work was also financed by the Higher Education Institutional Excellence Programme (FIKP) of the Ministry of Human Capacities in Hungary, within the framework of the Molecular Biology thematic programme of the Semmelweis University and by “Lendület” grants from the Hungarian Academy of Sciences to both MG (LP2011-001/2011) and BE (LP2018-13/2018). EB is also grateful for the support of the Premium postdoctoral research program (PPD-016/2017) of the Hungarian Academy of Sciences.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of Medicine, Department of PhysiologySemmelweis UniversityBudapestHungary
  2. 2.MTA-SE Lendﺲlet Tissue Damage Research GroupHungarian Academy of Sciences and Semmelweis UniversityBudapestHungary

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