The presence of ROS in the hippocampus, cortex and amygdala were probed using dihydroethidium (DHE) (Sigma Chemical Co). Brains were post-fixed, cryoprotected sequentially in 15 %, 30 % and 40 % sucrose, and embedded in tissue freezing medium for cryosectioning. Thick (25 μm) sagittal sections were prepared using a cryostat (Leica, Germany), mounted onto charged slides and then stored at -20℃ until staining. To test for the presence of ROS, sections from five non-cadmium treated HFD animals and five cadmium-treated HFD animals were incubated with 5 μM DHE at 37 °C for 30 min in the dark, as previously described [38 (link)]. To confirm the presence of superoxide, additional sections were pre-incubated for one hour at room temperature with Polyethylene Glycol-Conjugated Superoxide Dismutase (500 units/mL, Sigma) followed by DHE staining. Fluorescence images of hippocampus, cortex and amygdala regions of each animal were obtained using a laser scanning confocal microscope (Leica).
Polyethylene glycol conjugated superoxide dismutase
Manufactured by Merck Group
Sourced in United States
Polyethylene Glycol-Conjugated Superoxide Dismutase is a lab equipment product that serves as a conjugated form of the enzyme superoxide dismutase. It is used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Confocal laser scanning microscope, by Leica (1 mentions)
Dihydroethidium dhe, by Merck Group (1 mentions)
Cryostat, by Leica (1 mentions)
Diphenyleneiodonium chloride, by Merck Group (1 mentions)
Diethyldithiocarbamate, by Merck Group (1 mentions)
Diethylenetriaminepentaacetic acid, by Merck Group (1 mentions)
Catalase, by Merck Group (1 mentions)
Paraquat, by Thermo Fisher Scientific (1 mentions)
2 protocols using polyethylene glycol conjugated superoxide dismutase
1
Visualizing Oxidative Stress in Brain Regions
2
Cyclic Hydroxylamine Radical Detection
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Cyclic hydroxylamines 1-hydroxy-4-methoxy-2,2,6,6-tetramethylpiperidine (TM-H), and 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO) were purchased from Enzo Life Sciences (San Diego, CA, USA). Catalase, polyethylene glycol-conjugated superoxide dismutase (PEG-SOD), diethyldithiocarbamate (DDC), diethylenetriaminepenta-acetic acid (DTPA), diphenyleneiodonium chloride (DPI) and 4-Amino-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Paraquat was obtained from Fisher Scientific (Santa Clara, CA, USA). Stock solutions of cyclic hydroxylamine (10 mM) were prepared in argon-purged 0.9% NaCl treated with 0.1 mM DTPA. Stock solutions were prepared daily and kept in an ice bath under argon to avoid autooxidation. Certain commercial equipment, instruments, or materials are identified in this document. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the products identified are necessarily the best available for the purpose.
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