A simplified hydroethidine method for fast and accurate detection of superoxide production in isolated mitochondria

被引:15
|
作者
Back, Patricia [1 ]
Matthijssens, Filip [1 ]
Vanfleteren, Jacques R. [1 ]
Braeckman, Bart P. [1 ]
机构
[1] Univ Ghent, Dept Biol, Lab Aging Physiol & Mol Evolut, B-9000 Ghent, Belgium
关键词
Superoxide; Hydroethidine; C; elegans; Isolated mitochondria; AGING CAENORHABDITIS-ELEGANS; FLUORESCENCE DETECTION; OXIDATIVE STRESS; OXYGEN; PROBES; CELLS; EXPRESSION; ORGANISMS; TISSUES; ASSAY;
D O I
10.1016/j.ab.2012.01.008
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码:147 / 151
页数:5
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