Lipid peroxidation mda assay kit

The MDA content in tumor tissue lysates was measured using the Lipid Peroxidation MDA Assay Kit (Biosharp, BL904A) based on the reaction between MDA and thiobarbituric acid (TBA). Tumor tissues were homogenized using a tissue homogenizer, and the supernatant was collected by centrifugation at 12,000 × g for 10 min. Then 100 μl supernatant and 200 μl MDA detection working solution were mixed in a centrifuge tube. After being heated at 100 °C for 15 min, the supernatant was collected by centrifugation at 1000 × g at RT for 10 min. Totally, 200 μl of the reaction mix was transferred to a 96-well plate, and the absorbance at 532 nm was measured. The MDA standard curve was generated along the samples. Meantime, the protein concentration of supernatant from tissue lysate was determined after further dilution. The MDA concentration of each sample was calculated using the standard curve and normalized to the protein concentration.

Kidney tissue and cell samples were homogenized on ice using a homogenizer and then centrifuged for supernatant collection. Glutathione (GSH) levels were measured using a reduced GSH assay kit (Jiancheng, Nanjing, China), and optical density was measured at 405 nm. Malondialdehyde (MDA) levels were measured using a lipid peroxidation MDA assay kit (Biosharp, Hefei, China), and optical density was measured at 535 nm. Tissue iron levels were measured using a tissue iron assay kit (Jiancheng, Nanjing, China), and optical density was measured at 520 nm. Fe²⁺ concentrations were measured using a ferrous ion colorimetric assay kit (Elabscience, Wuhan, China), and optical density was measured at 590 nm. The kits were used following the manufacturers' instructions.