A simplified hydroethidine method for fast and accurate detection of superoxide production in isolated mitochondria.
Because superoxide is involved in various physiological processes, many efforts have been made to improve its accurate quantification. We optimized and validated a superoxide-specific and -sensitive detection method. The protocol is based on fluorescence detection of the superoxide-specific hydroethidine (HE) oxidation product, 2-hydroxyethidium. We established a method for the quantification of superoxide production in isolated mitochondria without the need for acetone extraction and purification chromatography as described in previous studies.
Laboratory for Aging Physiology and Molecular Evolution, Department of Biology, Ghent University, 9000 Ghent, Belgium.
Patricia
P
Back
Filip
F
Matthijssens
Jacques R
JR
Vanfleteren
Bart P
BP
Braeckman
D000096:Acetone / Q000737:chemistry; D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D017173:Caenorhabditis elegans; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D004996:Ethidium / Q000031:analogs & derivatives / Q000032:analysis; D005470:Fluorometry; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D010084:Oxidation-Reduction; D010617:Phenanthridines / Q000737:chemistry*; D013481:Superoxides / Q000032:analysis*
C506392:2-hydroxyethidium; D010617:Phenanthridines; D013481:Superoxides; D000096:Acetone; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; C058428:hydroethidine; D000968:Antimycin A; D004996:Ethidium