To evaluate the occurrence of ferroptosis in heart tissue, freshly isolated heart tissue was lysed to prepare tissue homogenates. Malondialdehyde (MDA) levels in heart tissues and H9c2 cells were measured by using a commercial kit (S0131; Beyotime, China) in accordance with the manufacturer’s instructions. Ferrous ion levels in heart tissues was quantified using an Iron Assay Kit (BC4355, Solarbio, Beijing) and those in H9c2 cells was quantified using an Iron Assay Kit (E1042, Alphabio, Tianjin) according to the manufacturer's instructions. The nonenzymatic antioxidant system in heart tissues and H9c2 cells was assessed by examining glutathione (GSH) and oxidized glutathione (GSSG) levels and the GSH/GSSG ratio using a GSH and GSSG Assay Kit (A061-1, Nanjing Jiancheng BioTech, China) according to the manufacturer's instructions.
Gsh and gssg assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The GSH and GSSG Assay Kit is a laboratory equipment product designed to measure the levels of glutathione (GSH) and glutathione disulfide (GSSG) in various biological samples. It provides a quantitative analysis of these important antioxidant molecules, which play a crucial role in cellular redox homeostasis and oxidative stress response.
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2 protocols using gsh and gssg assay kit
1
Ferroptosis Evaluation in Heart Tissue
2
Intracellular ROS and Oxidative Stress Assays
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Intracellular reactive oxygen species (ROS) was examined by a peroxide‐sensitive fluorescent probe DCFH‐DA (S0033S, Beyotime). A549 and H1975 cells were cultured at 37°C for 1 h with 10 μM of DCFH‐DA, and the fluorescence intensity was calculated with a fluorescence microscope. ROS level (%) = fluorescence value of intervention group/control group × 100%. In the tumor tissues, DCFH‐DA was added to the lysed tissues and incubated for 20 min at 37°C, washed three times with phosphate buffered saline (PBS), and the fluorescence intensity was detected.
Malondialdehyde (MDA) content (A003‐1‐2, Nanjing Jiancheng) and superoxide dismutase (SOD) activity (A001‐3‐2, Nanjing Jiancheng) were measured with the relevant kits. Collected A549 and H1975 cells were sonicated and tumor tissues were homogenized. MDA content and SOD activity were evaluated according to cell and tissue protein concentrations.
Total glutathione (GSH) content was assayed by GSH and GSSG assay kit (A061‐2‐1, Nanjing Jiancheng) and evaluated by comparison with the GSH standard curve.
Malondialdehyde (MDA) content (A003‐1‐2, Nanjing Jiancheng) and superoxide dismutase (SOD) activity (A001‐3‐2, Nanjing Jiancheng) were measured with the relevant kits. Collected A549 and H1975 cells were sonicated and tumor tissues were homogenized. MDA content and SOD activity were evaluated according to cell and tissue protein concentrations.
Total glutathione (GSH) content was assayed by GSH and GSSG assay kit (A061‐2‐1, Nanjing Jiancheng) and evaluated by comparison with the GSH standard curve.
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