AAT Bioquest

What methods can I use to measure oxidative stress?

Posted June 30, 2023


There are several methods that can be used to measure oxidative stress. One method is using the DCFH-DA assay to directly measure the intracellular levels of reactive oxygen species. DCFH-DA (2,7-dichlorodihydrofluorescein diacetate) is oxidized by ROS to create the fluorescent compound 2,7-dichlorofluorescin. This compound can be detected by utilizing flow cytometry or fluorescence microscopy. A hydrogen peroxide assay may also be used to measure oxidative stress, and use peroxide-sensitive probes in its procedure. Protein carbonylation assays are also markers of oxidative damage to protein. Carbonyl groups of oxidized proteins are exposed to DNPH and the resulting protein-DNPH can be quantified using spectrophotometric techniques. Lastly, oxidative stress can be measured indirectly by measuring the levels of DNA/RNA damage, rather than directly measuring the ROS. It can be quantified using chromatography or ELISA assays. 

Additional resources

Approaches and Methods to Measure Oxidative Stress in Clinical Samples: Research Applications in the Cancer Field

Reactive Oxygen Species (ROS)

ROS Brite™ 570 *Optimized for Detecting Reactive Oxygen Species (ROS)*

Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Orange Fluorescence*