Mitochondrial ROS emission was measured as previously described with minor modifications (56 (link)). Briefly, mitochondria were incubated in a black 96-well plate with mitochondrial respiration buffer and 5 μm Amplex Red (Thermo Fisher Scientific, Waltham, MA, USA), 1 U/ml Horseradish peroxidase (169-10791, Fujifilm Wako Pure Chemical Corp.), and 5 U/ml superoxide dismutase (192-11281, Fujifilm Wako Pure Chemical Corp.). The kinetics of ROS emission were assessed under state 3 respiratory conditions through the addition of 2.5 mm ADP, 10 mm glutamate and 2 mm malate. The relative changes in fluorescence per minute were measured using a fluorescence plate reader (Spark 20 M, Tecan) (Ex: 560 [20] nm; Em: 620 [20] nm). The relative change in fluorescence per minute was measured and normalized to OCR.
Superoxide dismutase
Manufactured by Fujifilm
Sourced in United States, Japan
Superoxide dismutase is a laboratory reagent that catalyzes the dismutation of superoxide radicals into oxygen and hydrogen peroxide. It is an antioxidant enzyme that plays a vital role in protecting cells from oxidative stress.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using superoxide dismutase
1
Mitochondrial ROS Emission Measurement
2
Antioxidant Activity Evaluation Protocol
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Linseed oil, corn oil, H2O2 (30%), K3[Fe(CN)6], catalase (CAT: from bovine liver, 10000 U/mg) and superoxide dismutase (SOD: from bovine erythrocyte, 3500 U/mg) were obtained from Wako Pure Chemicals Co. (Osaka, Japan). Canola oil, its preparations (A and B) for O/W emulsion (A and B), and sucrose stearic acid ester (RYOTO TM Sugar Ester S-1170; HLB 11) emulsifier were obtained from Mitsubishi-Kagaku Foods Co. (Kanagawa, Japan). Preparation A consisted of 85% canola oil and 15% emulsifiers, and preparation B was 40% canola oil, 55% sugar-alcohol and 5% emulsifiers. TBA, 1,1,3,3-tetraethoxypropane (TEP) and cumen hydroperoxide (CHP) were obtained from Tokyo Kasei Kogyo Co. (Tokyo, Japan). Luminol and Triton X-100 were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Tween 20 and FeCl2•4H2O were purchased from Nacalai Tesque (Tokyo, Japan). Distilled water was passed through a Pure Line WL21P system (Yamato Scientific Co., Tokyo, Japan). All other chemicals were of analytical grade. A standard stock solution of malondialdehyde (5 mM) for the TBA assay was prepared by dissolving TEP in H2O and storing at 4°C until analysis. 15
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