ROS Detection Cell-Based Assay Kit (DHE)

Dihydroethidium (hydroethidine or DHE) is a widely used ethidium-based, redox-sensitive fluorescent probe. 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).^1,2 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. This assay kit uses DHE as a fluorescent probe for the detection of ROS generation. 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 antioxidant control.

Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.

WARNING This product is not for human or veterinary use.

1. Michalski, R., Michalowski, B., Sikora, A., et al. On the use of fluorescence lifetime imaging and dihydroethidium to detect superoxide in intact animals and ex vivo tissues: A reassessment. Free Redic. Biol. Med. 67, 278-284 (2014).

2. Zhao, H., Kalivendi, S., Zhang, H., et al. Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: Potential implications in intracellular fluorescence detection of superoxide. Free Radic. Biol. Med. 34(11), 1359-1368 (2003).

Bohovych, I., Fernandez, M.R., Rahn, J.J., et al. Metalloprotease OMA1 fine-tunes mitochondrial bioenergetic function and respiratory supercomplex stability. Sci. Rep. 5, 13989 (2015).

Zhang, J., Cruz-Cosme, R., Zhang, C., et al. Endoplasmic reticulum-associated SARS-CoV-2 ORF3a elicits heightened cytopathic effects despite robust ER-associated degradation. mBio 15(1), e0303023 (2026).