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Hydroethidine is a fluorogenic probe that in the presence of the superoxide radical anion is oxidized to a red fluorescent product, 2-hydroxyethidium. In cells, hydroethidine is also oxidized to other products, including red fluorescent ethidium. Thus, selective monitoring of 2-hydroxyethidium is required for specific detection of the superoxide radical anion. Here, we provide protocols for HPLC- and LC-MS-based quantitation of 2-hydroxyethidium, among other oxidation products. Also, a protocol for continuous sampling for real-time monitoring of superoxide production using rapid HPLC measurements of 2-hydroxyethidium is described.
Pubmed ID: 31172476
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Redox and Bioenergetics core provides instrumentation, cutting-edge techniques, and expertise dedicated to investigating cancer cell metabolism and redox signaling, and to design custom probes for reactive oxygen species (ROS) for MCWCC members.
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