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@ARTICLE{Eid:303468,
      author       = {M. Eid$^*$ and U. Barayeu$^*$ and T. Dick$^*$},
      title        = {{C}hemogenetic detection and quantitation of {H}2{O}2 in
                      living cells.},
      journal      = {Nature protocols},
      volume       = {nn},
      issn         = {1754-2189},
      address      = {Basingstoke},
      publisher    = {Nature Publishing Group},
      reportid     = {DKFZ-2025-01665},
      pages        = {nn},
      year         = {2025},
      note         = {DKFZ-ZMBH Alliance / #EA:A160#LA:A160#},
      abstract     = {Hydrogen peroxide (H2O2) is a natural product of aerobic
                      metabolism. It acts as a signaling molecule and regulates
                      fundamental cellular functions. However, it has remained
                      difficult to measure intracellular H2O2 with high
                      specificity and in a quantitative manner. Here, we present a
                      detailed protocol for a chemogenetic method that enables the
                      detection and quantitation of H2O2 in living cells by
                      converting intracellular H2O2 into fluorescent or
                      luminescent signals. This is achieved by expressing the
                      engineered heme peroxidase APEX2 in cells and subcellular
                      locations of interest and by providing an appropriate
                      fluorogenic or luminogenic substrate from outside. This
                      method differs fundamentally from previously developed
                      genetically encoded H2O2 probes; those are reversible and
                      measure the balance between probe thiol oxidation and
                      reduction. By contrast, APEX2 turns over its substrate
                      irreversibly and therefore directly measures endogenous H2O2
                      availability. Our detailed step-by-step protocol covers the
                      generation of APEX2-expressing cell lines, the
                      implementation of fluorescent and luminescent measurements
                      and examples for application. Ectopic expression of APEX2
                      can be achieved in 3 days, while the actual measurements
                      typically require 1-2 h. This protocol is intended for
                      entry-level scientists.},
      subtyp        = {Review Article},
      cin          = {A160},
      ddc          = {610},
      cid          = {I:(DE-He78)A160-20160331},
      pnm          = {311 - Zellbiologie und Tumorbiologie (POF4-311)},
      pid          = {G:(DE-HGF)POF4-311},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:40790259},
      doi          = {10.1038/s41596-025-01226-9},
      url          = {https://inrepo02.dkfz.de/record/303468},
}