Malondialdehyde mda content assay kit

The H₂O₂ content of samples was determined using a Hydrogen Peroxide Assay Kit (Solaibao Technology, Beijing, China) according to the manufacturer’s instructions. In total, 0.1 g leaf samples with or without the 140 mM NaCl treatment were mixed with 1 mL of propanone and centrifuged at 8000× g for 10 min at 4 °C. Further, 250 μL of supernatant and 325 μL of test solution (5% titanic sulfate) were mixed and placed at 25 °C for 10 min. The absorbance was immediately measured at 415 nm using a spectrometer. The H₂O₂ content in the sample was calculated using its calibration curve. The MDA content was determined using a Malondialdehyde (MDA) content Assay Kit from Solarbio Technology (Beijing, China). Rice leaves were homogenized in 1 mL of 10% trichloroacetic acid (TCA) and centrifuged at 8000× g for 10 min at 4 °C. Further, 100 μL of the supernatant was mixed with 200 μL of 0.67% thiobarbituric acid (TBA) and placed in a boiling water bath for 60 min. Further, the mixture was centrifuged at 4000× g and 4 °C for 10 min, and the absorbance of the supernatant was measured at 450, 532, and 600 nm.

The levels of hydrogen peroxide (H₂O₂), malondialdehyde (MDA), and chlorophyll in cotton seedling leaves were determined using the Hydrogen Peroxide Content Assay Kit (Solarbio, Beijing, China), Malondialdehyde (MDA) Content Assay Kit (Solarbio), and Chlorophyll Assay Kit (Solarbio) according to the manufacturer’s instructions, respectively. All the analyses were carried out in triplicate.

All following drugs and reagents were acquired commercially: Valsartan (Sigma-Aldrich, St. Louis, MO, United States); Lipopolysaccharide (Sigma-Aldrich, St. Louis, MO, United States); Cell Counting Kit-8 (CCK-8) (Biosharp, Guangzhou, China); Malondialdehyde (MDA) Content Assay Kit (Solarbio, Beijing, China); Oxidized Glutathione (GSSG) Content Assay Kit (Solarbio, Beijing, China); Superoxide Dismutase (SOD) Activity Assay Kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China); Antibodies against p-AKT, AKT, P38, p-JNK, JNK, ERK and MUC5AC (Zen-bio, Chengdu, China). Antibodies against p-P38, p-ERK, and Tubulin (Affinity Biosciences, Jiangsu, China). Antibodies against GAPDH and β-actin (ABclonal, Wuhan, China).

The differences in the physiological responses of the ‘268’ and ‘334’ lines under control and heat stress treatments were measured during CK, HT-4, HT-8, HT-10, and RC-4 periods. In accordance with the manufacturer’s instructions, the corresponding kits (Solarbio Life Sciences, Beijing, China) were selected to determine various physiological indicators. The Catalase (CAT) Activity Assay Kit (Cat#BC0200; Solarbio, Beijing, China), Superoxide Dismutase (SOD) Activity Assay Kit (Cat#BC0170; Solarbio, Beijing, China), Proline (Pro) Content Assay Kit (Cat#BC0290; Solarbio, Beijing, China), Malondialdehyde (MDA) Content Assay Kit (Cat#BC0020; Solarbio, Beijing, China), and Plant Soluble Sugar (SS) Content Assay Kit (Cat#BC0030; Solarbio, Beijing, China) were used for subsequent spectrophotometric measurements. Three biological replicates of each sample (n = 3) were assessed.