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JoVE Journal Biochemistry
Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
 

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases

Article DOI: 10.3791/56975-v • 08:57 min • February 24th, 2018
February 24th, 2018 •
1, 1, 1, 1
1Department of Biochemistry, Memorial University of Newfoundland

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Mitochondria contain several flavin-dependent enzymes that can produce reactive oxygen species (ROS). Monitoring ROS release from individual sites in mitochondria is challenging due to unwanted side reactions. We present an easy, inexpensive method for direct assessment of native rates for ROS release using purified flavoenzymes and microplate fluorometry.

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Superoxide Hydrogen Peroxide NADH Flavin-containing Mitochondrial Dehydrogenases Microplate Fluorometry Production Rate Substrate Inhibitor Combinations Isolated Mitochondria Release Capacity Reactive Oxygen Species Antioxidants Specificity Additives Simultaneous Measurement
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