DHE (Dihydroethidium) Assay Kit - Reactive Oxygen Species

DHE (Dihydroethidium) Assay Kit - Reactive Oxygen Species (ab236206) measures ROS directly in live cells. This kit uses DHE as a fluorescent probe for the detection of ROS generation and is specific for superoxide and hydrogen peroxide. Antimycin A, an inhibitor of complex III of the mitochondrial electron transport chain, is included as a positive control for ROS generation. N-acetyl Cysteine is included as an anti-oxidant control.

ROS generation is represented as total DHE fluorescence. It is important that positive and negative controls be included in every experiment for all cell types tested. Data can be expressed as total fluorescence, geometric mean fluorescence (flow cytometry) or as % or controls.

DHE has been shown to be oxidized by superoxide to form 2-hydroxyethidium (2-OH-E⁺) (ex 500-530 nm/em 590-620 nm) or by non-specific oxidation by other sources of reactive oxygen species (ROS) to form ethidium (E⁺) (ex 480 nm/em 576 nm). Given their narrow spectral range, distinguishing between the two species using filter-based optical systems is often difficult and can lead to misreporting of the species of ROS being generated.