Analyses of mitochondrial ROS production

HL Hakjoo Lee
TL Tae Jin Lee
CG Chad A. Galloway
WZ Wenbo Zhi
WX Wei Xiao
KB Karen L. de Mesy Bentley
AS Ashok Sharma
YT Yong Teng
HS Hiromi Sesaki
YY Yisang Yoon
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Mitochondrial ROS production was analyzed by using the Amplex Red H2O2 assay kit (Invitrogen) per manufacturer’s instruction. Mitochondria (250 μg) was added to the reaction mixture containing Amplex Red (50 μM), horse radish peroxidase (0.1 U/ml), superoxide dismutase (80 U/ml), and 5 mM glutamate / 2.5 mM malate or 5 mM succinate, and the fluorescence was measured at 590 nm with 540 nm excitation (APD130A2 Avalanche Photodetector, Thorlabs Inc.).

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