The specific probe hydroethidine (Thermo Fisher Scientific, Grand Island, NY, USA) was used to measure changes in cellular superoxide (O2•−) production[59 (link)]. The reaction between O2•− and non-fluorescent hydroethidine generates a highly specific red fluorescent product 2-hydroxyethidium. In biological systems, another red fluorescent product ethidium is also formed, usually at a much higher concentration than 2-hydroxyethidium[60 (link)]. Cultured cells, grown on glass slides (4 independent experiments per group), were incubated with hydroethidine (2 μg/ml) at room temperature for 15–20 min and then fixed with 4% paraformaldehyde. Increased O2•− production was quantified at 488 nm/ > 510 nm (excitation/emission). For confocal microscopy studies, the cultured cells were fixed with 4% paraformaldehyde at room temperature for 15–20 min and stored in phosphate buffer saline at 4
Specific probe hydroethidine
Manufactured by Thermo Fisher Scientific
Sourced in United States
Specific probe hydroethidine is a fluorescent probe used for the detection and quantification of superoxide anion (O2-) in biological samples. It functions by reacting with superoxide to produce a fluorescent product, enabling the visualization and measurement of superoxide levels in cells or tissues.
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Lab products found in correlation
2 protocols using specific probe hydroethidine
1
Measuring Cellular Superoxide Production
2
Measuring Cellular Superoxide Production
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The specific probe hydroethidine (Thermo Fisher Scientific, Waltham, MA, United States, cat # D11347) was used to measure changes in cellular superoxide (O2•–) production (Sen and Hongpaisan, 2018 (link)). The reaction between O2•– and non-fluorescent hydroethidine generates a highly specific red fluorescent product, 2-hydroxyethidium. In biological systems, another red fluorescent product ethidium is also formed, usually at a much higher concentration than 2-hydroxyethidium (Zielonka and Kalyanaraman, 2010 (link)). Cultured cells, grown on glass slides, were incubated with hydroethidine (2 μg/ml) at room temperature for 15–20 min and then fixed with 4% paraformaldehyde. Increased O2•– production was qualitatively determined with a confocal microscope at 488 nm/>510 nm (excitation/emission).
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