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Related Concept Videos

Redox Reactions01:27

Redox Reactions

Redox reactions are vital biochemical processes that underpin energy metabolism in cells. These reactions involve the transfer of electrons between molecules, occurring in tandem as oxidation and reduction. Oxidation refers to the loss of electrons, while reduction denotes their gain. This coupling ensures the seamless flow of electrons through metabolic pathways. For example, in bacterial metabolism, glucose undergoes oxidation to carbon dioxide, while oxygen is simultaneously reduced to...
Redox Reactions01:24

Redox Reactions

Oxidation-reduction or redox reactions involve the transfer of electrons from one molecule or atom to another. When an atom gains an electron, another atom must lose an electron, meaning oxidation and reduction must occur together. Since the redox occurs in pairs, the atom that gets oxidized is also called the reducing agent or reductant, and the atom that is reduced is also called the oxidizing agent or oxidant. A straightforward way to remember the definitions of oxidation and reduction is...
Redox Titration: Other Oxidizing and Reducing Agents01:26

Redox Titration: Other Oxidizing and Reducing Agents

Besides iodine, other oxidizing or reducing agents can serve as titrants in redox titrations. Common oxidizing titrants include KMnO4, cerium(IV), and K2Cr2O7. The choice of oxidizing titrants depends on factors like stability, cost, analyte strength, and reaction rate between the analyte and titrant. KMnO4 is a strong oxidizing titrant that reduces from Mn(VII) to Mn(II) in a highly acidic solution, simultaneously oxidizing the analyte to a higher oxidation state. In this case, KMnO4 acts as a...
Redox Equilibria: Overview01:23

Redox Equilibria: Overview

A reduction-oxidation reaction is commonly called a redox reaction. In a redox reaction, electrons are transferred from one species to another rather than being shared between or among atoms. The reducing agent or reductant is the species that loses electrons and gets oxidized in the process. The species that gains electrons and gets reduced in the process is the oxidizing agent or oxidant. Redox reactions are represented as two separate equations called half-reactions, where one equation...
Redox Titration: Overview01:21

Redox Titration: Overview

Redox titration is a chemical analysis technique used to determine the concentration of an unknown substance by measuring the electron transfer in a redox (reduction-oxidation) reaction. The process involves gradually adding a titrant with a known concentration of an oxidizing or reducing agent, to the analyte, the solution with an unknown concentration, until reaching the endpoint, which indicates the completion of the reaction between the two substances. Ensuring the analyte is in a single...

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Related Experiment Video

Updated: May 28, 2026

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment
10:46

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

Published on: March 16, 2018

Measuring E(GSH) and H2O2 with roGFP2-based redox probes.

Bruce Morgan1, Mirko C Sobotta, Tobias P Dick

  • 1Division of Redox Regulation, DKFZ–ZMBH Alliance, German Cancer Research Center, Heidelberg, Germany

Free Radical Biology & Medicine
|October 4, 2011
PubMed
Summary

This study introduces new tools for measuring redox states in live cells. The tools are genetically encoded biosensors based on roGFP2. They can track glutathione redox potential and hydrogen peroxide levels. The methods work in yeast and mammalian cells. The probes allow precise, real-time measurements. The study provides detailed protocols for using the biosensors. These tools improve the ability to study redox biology. They help differentiate between different redox signals. The findings support using genetically encoded probes in redox research.

Keywords:
redox biosensorsgenetically encoded probescellular redox statefluorescent redox measurement

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Area of Science:

  • Redox biochemistry in cell biology
  • Fluorescent biosensor development in molecular imaging
  • Genetically encoded probe applications in biotechnology

Background:

Redox processes influence many cellular functions. Yet, measuring redox states remains challenging. Multiple redox couples exist, and they do not always equilibrate. This complicates interpretations of overall cellular redox status. Different systems may carry distinct biological signals. A general "cellular redox state" is not meaningful for all contexts. Prior research has shown that redox couples vary by compartment and cell type. That uncertainty drove the need for precise, localized measurements. No prior work had resolved how to track specific redox couples in real time. This gap motivated the development of genetically encoded redox biosensors.

Purpose Of The Study:

This study aims to improve redox measurements in living cells. It focuses on specific redox couples rather than global states. The goal is to enable precise, in vivo measurements. The authors propose using genetically encoded roGFP-based probes. These tools allow tracking of E(GSH) and H2O2 levels. The study provides protocols for yeast and mammalian systems. It emphasizes spatiotemporal resolution in redox monitoring. The researchers propose that these probes can advance redox biology research.

Main Methods:

The study uses genetically encoded roGFP2-based biosensors. These probes are fused to redox-active proteins. Grx1-roGFP2 targets glutathione redox potential. roGFP2-Orp1 measures hydrogen peroxide changes. The methods include yeast and mammalian cell systems. Plate-reader and microscopy techniques are described. Rapid equilibration of roGFP with target redox couples is ensured. The protocol outlines steps for probe expression and measurement.

Main Results:

The Grx1-roGFP2 probe effectively measures E(GSH) in cells. The roGFP2-Orp1 probe tracks H2O2 concentration changes. Both probes show rapid equilibration with their targets. The methods work in both yeast and mammalian cells. Plate-reader and microscopy approaches are validated. The probes enable redox couple-specific measurements. The results suggest these tools can provide detailed redox data. The authors report that these methods improve spatial and temporal resolution.

Conclusions:

The authors propose that roGFP-based probes advance redox biology. These tools allow specific redox couple measurements in live cells. The probes work in yeast and mammalian systems. The study shows that E(GSH) and H2O2 can be tracked effectively. The methods improve spatiotemporal resolution of redox data. The authors suggest that these probes can replace less specific techniques. They propose that these tools can help differentiate redox signals. The findings support the use of genetically encoded biosensors in redox research.

The probes measure glutathione redox potential (E(GSH)) and hydrogen peroxide (H2O2) levels in live cells.

The probes are genetically fused to redox-active proteins, allowing rapid equilibration with specific redox couples.

E(GSH) reflects the redox state of glutathione, a key antioxidant in cellular defense mechanisms.

Yes, the study provides protocols for using the probes in both yeast and mammalian systems.

The study uses plate-reader and microscopy-based methods to measure fluorescence changes.

The probes allow redox couple-specific measurements with high spatial and temporal resolution.