Experimental workflows for the accurate identification of mitochondrial redox events

@article{hernansanzagustin2026experimental,
  title = {{Experimental workflows for the accurate identification of mitochondrial redox events}},
  author = {Pablo Hernansanz-Agustín and Juan Pedro Bolaños and Consuelo Borrás and Elisa Cabiscol and Victoria Campuzano and Laura de Cubas and José Antonio Enríquez and Isabel Fabregat and Gemma Gonfaus-Ortiz and Elena Hidalgo and Marc Liesa and Cristina Mas-Bargues and Antonio Martínez-Ruiz and María Monsalve and Rubén Quintana-Cabrera and Sergio Rius-Pérez and Juan Sastre and Antonio Zorzano},
  journal = {Redox Biology},
  year = {2026},
  month = oct,
  doi = {10.1016/j.redox.2026.104387},
  url = {https://doi.org/10.1016/j.redox.2026.104387},
  note = {CC BY},
}

TY  - JOUR
TI  - Experimental workflows for the accurate identification of mitochondrial redox events
AU  - Pablo Hernansanz-Agustín
AU  - Juan Pedro Bolaños
AU  - Consuelo Borrás
AU  - Elisa Cabiscol
AU  - Victoria Campuzano
AU  - Laura de Cubas
AU  - José Antonio Enríquez
AU  - Isabel Fabregat
AU  - Gemma Gonfaus-Ortiz
AU  - Elena Hidalgo
AU  - Marc Liesa
AU  - Cristina Mas-Bargues
AU  - Antonio Martínez-Ruiz
AU  - María Monsalve
AU  - Rubén Quintana-Cabrera
AU  - Sergio Rius-Pérez
AU  - Juan Sastre
AU  - Antonio Zorzano
PY  - 2026
DA  - 2026/10/30
JO  - Redox Biology
DO  - 10.1016/j.redox.2026.104387
UR  - https://doi.org/10.1016/j.redox.2026.104387
AB  - The study of redox biology has been growing constantly since the last decades. Over these years, redox processes have been linked to an extraordinarily wide range of physiological and pathological events, becoming recognized as central mechanisms underlying many of them. In this context, it becomes essential to understand the advantages and limitations of the tools under use, to recognize the specific controls required for each measurement and to accurately distinguish between distinct redox mechanisms. So far, multiple and excellent reviews have dealt with either the tools, the protocols or the mechanisms involved in reactive oxygen species (ROS) production and quenching, a.k.a. redox events. However, a review outlining the workflows to appropriately detect them is still lacking. We define workflow as the combination of tools, methods and mechanistic knowledge that allow the definition of a specific redox event. In this review, we aim to provide an optimal workflow for the research on mitochondrial redox events. To this end, we first summarize the molecular tools available to measure and quench ROS. We then explain the mechanisms of ROS production and scavenging in several of the cellular compartments, with special focus on mitochondria, as well as their implication in physiology and disease. Finally, we use the knowledge in all sections to build a recommended experimental workflow, illustrated by several cases of study. This review will enable the reader to understand how specific mitochondrial redox events can be accurately measured, considering all technical, methodological and mechanistical variables and limitations required for their reliable detection and interpretation.
SN  - 2213-2317
ER  -