ROS Detection Cell-Based Assay Kit (DHE) Usage And Synthesis

Description

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). 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.

References

[1] RADOSLAW MICHALSKI . On the use of fluorescence lifetime imaging and dihydroethidium to detect superoxide in intact animals and ex vivo tissues: A reassessment[J]. Free Radical Biology and Medicine, 2014, 67: Pages 278-284. DOI: 10.1016/j.freeradbiomed.2013.10.816
[2] HONGTAO ZHAO . Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide[J]. Free Radical Biology and Medicine, 2003, 34 11: Pages 1359-1368. DOI: 10.1016/s0891-5849(03)00142-4
[3] IRYNA BOHOVYCH. Metalloprotease OMA1 Fine-tunes Mitochondrial Bioenergetic Function and Respiratory Supercomplex Stability.[J]. Scientific Reports, 2015: 13989. DOI: 10.1038/srep13989

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