The oxidative stress in the brain at 24 hours after CA was evaluated by immunohistochemical staining of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Brain tissues were sectioned and embedded in paraffin. The immunohistochemistry was performed as we previously described (28 (link)), and 8-OHdG antibody (diluted 1:200; Abcam, Cambridge, UK) was used. The immunostaining scores were estimated using both the percentage of positively stained cells and the staining intensity as we previously described (17 (link)). The ROS levels in the brain at 6 hours after CA were measured using the tissue ROS assay kit (Genmed Scientifics, Wilmington, DE, USA) that utilized 2’,7’-dichlorofluorescein diacetate as the oxidative fluorescent probe. After the fresh brain tissues were isolated, the ROS levels were measured following the manufacturer’s instructions.
Tissue ros assay kit
Manufactured by Genmed Scientifics
Sourced in United States
The Tissue ROS Assay Kit is designed to measure reactive oxygen species (ROS) levels in tissue samples. It provides a quantitative assessment of oxidative stress by detecting the presence and concentration of various ROS, including superoxide, hydrogen peroxide, and hydroxyl radicals. The kit utilizes a fluorescent probe that reacts with ROS, allowing for the determination of ROS levels through fluorescence measurement.
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2 protocols using tissue ros assay kit
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Oxidative Stress Evaluation in Brain after Cardiac Arrest
2
Oxidative Stress Evaluation After Cardiac Arrest
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The oxidative stress in the heart tissue at 24 h after CA was evaluated by immunohistochemical staining of 4-hydroxynonenal (4-HNE) to assess lipid peroxidation and 8-hydroxy-2′-deoxyguanosine (8-OHdG) to detect the extent of nucleic acid oxidation. Heart tissues were sectioned and embedded in paraffin. For immunohistochemistry, the fixed sections were immunostained overnight at 4°C using a primary monoclonal antibody against 4-HNE and 8-OHdG (Abcam, Cambridge, United Kingdom), followed by corresponding secondary antibody for 2.5 h at room temperature. Ten fields of vision (magnification of ×400) were randomly sampled from each heart tissue sample under an optical microscope. 4-HNE and 8-OHdG relative intensity area (%) were detected by automated counting software (Image J, National Institute of Health, United States). The ROS levels in the heart at 6 h after CA were measured using a tissue ROS assay kit (Genmed Scientifics, Wilmington, DE, United States) according to the manufacturer’s instructions.
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