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

来源：

ANALYTICAL BIOCHEMISTRY | 2012年 / 423卷 / 01期

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

页码：147 / 151

页数：5

共 23 条

[1] Effects of sod gene overexpression and deletion mutation on the expression profiles of reporter genes of major detoxification pathways in Caenorhabditis elegans [J].

Back, Patricia ;

Matthijssens, Filip ;

Vlaeminck, Caroline ;

Braeckman, Bart P. ;

Vanfleteren, Jacques R. .

EXPERIMENTAL GERONTOLOGY, 2010, 45 (7-8) :603-610

[2] Use of spectroscopic probes for detection of reactive oxygen species [J].

Bartosz, Grzegorz .

CLINICA CHIMICA ACTA, 2006, 368 (1-2) :53-76

[3]

Bindokas VP, 1996, J NEUROSCI, V16, P1324

[4] Assaying metabolic activity in ageing Caenorhabditis elegans [J].

Braeckman, BP ;

Houthoofd, K ;

De Vreese, A ;

Vanfleteren, JR .

MECHANISMS OF AGEING AND DEVELOPMENT, 2002, 123 (2-3) :105-119

[5] Disruption of insulin signalling preserves bioenergetic competence of mitochondria in ageing Caenorhabditis elegans [J].

Brys, Kristel ;

Castelein, Natascha ;

Matthijssens, Filip ;

Vanfleteren, Jacques R. ;

Braeckman, Bart P. .

BMC BIOLOGY, 2010, 8

[6] Against the oxidative damage theory of aging: superoxide dismutases protect against oxidative stress but have little or no effect on life span in Caenorhabditis elegans [J].

Doonan, Ryan ;

McElwee, Joshua J. ;

Matthijssens, Filip ;

Walker, Glenda A. ;

Houthoofd, Koen ;

Back, Patricia ;

Matscheski, Andrea ;

Vanfleteren, Jacques R. ;

Gems, David .

GENES & DEVELOPMENT, 2008, 22 (23) :3236-3241

[7] Oxidants, oxidative stress and the biology of ageing [J].

Finkel, T ;

Holbrook, NJ .

NATURE, 2000, 408 (6809) :239-247

[8] An ultrasensitive fluorescent assay for the in vivo quantification of superoxide radical in organisms [J].

Georgiou, CD ;

Papapostolou, I ;

Patsoukis, N ;

Tsegenidis, T ;

Sideris, T .

ANALYTICAL BIOCHEMISTRY, 2005, 347 (01) :144-151

[9] Superoxide radical detection in cells, tissues, organisms (animals, plants, insects, microorganisms) and soils [J].

Georgiou, Christos D. ;

Papapostolou, Ioannis ;

Grintzalis, Konstantinos .

NATURE PROTOCOLS, 2008, 3 (11) :1679-1692

[10] Fluorescence probes used for detection of reactive oxygen species [J].

Gomes, A ;

Fernandes, E ;

Lima, JLFC .

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 2005, 65 (2-3) :45-80

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