Superoxide dismutase sod assay kit

Defatted C. moschata seed meal was obtained from Anyang Jingsen Biotechnology Co., Ltd. (Henan, China). Alcalase (200 U/mg) and trypsin (250 U/mg) were obtained from Solarbio (Beijing, China). Dulbecco's Modified Eagle Medium (DMEM), streptomycin, and fetal bovine serum (FBS) were obtained from BNCC (Beijing, China). Hydrogen peroxide (H₂O₂) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma Biochemicals (St. Louis, MO, USA). 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), superoxide dismutase (SOD) assay kit, and GSH assay kit were purchased from Beyotime (Nanjing, China). Other chemicals used in the study were of analytical grade.

Fisetin, glucose and dimethyl sulfoxide (DMSO) were obtained from Sigma–Aldrich (St. Louis, MO, United States). Fisetin (the purity is > 98%) was dissolved in < 0.1% of DMSO solution. LY294002 (the specific inhibitor of PI3K) was bought from MedChemExpress (Shanghai, China). Cell counting kit-8 (CCK-8) was purchased from Dojindo China Co., Ltd. (Shanghai, China). LDH cytotoxicity assay kit, lipid peroxidation [malondialdehyde (MDA)] assay kit, superoxide dismutase (SOD) assay kit, and Hoechst 33258 were obtained from Beyotime (Shanghai, China). Dulbecco’s modified eagle’s medium (DMEM) and phosphate-buffered saline (PBS) were purchased from Biological Industries (Shanghai, China). Fetal bovine serum (FBS) was purchased from Gibco (Grand Island, NY, United States). Antibody against PI3K (#4257), Phospho-PI3Kp85/p55 (#4228), Akt (#4691), Phospho-Akt (#4060), CREB (#4820), and p-CREB (#9198) were obtained from Cell Signaling Technology (Beverly, MA, United States). Antibody against β-actin was purchase from Absin (Shanghai, China).

ICA (>98% purity) was purchased from Nanjing Zelang Pharmaceutical Technology (Nanjing, China). DMSO was purchased from Sigma. The standard MSC expansion medium, osteogenesis kit, adipogenesis kit, and chondrogenic kit were purchased from Cyagen Biosciences Inc. (Guangzhou, China). Cell counting kit-8 (CCK-8) was purchased from Dojindo (Japan). TUNEL staining was purchased from Roche Diagnostics GmbH (Roche, Germany). Phalloidin conjugated with Alexa Fluor 488 and DAPI was purchased from Sigma. Annexin V-FITC/PI apoptosis detection kit, lactate dehydrogenase- (LDH-) cytotoxicity assay kit, JC-1 staining, ROS detection kit, glutathione peroxidase (GPx) assay kit, superoxide dismutase (SOD) assay kit, lipid peroxidation malondialdehyde (MDA) assay kit, and western and IP cell lysis kit were purchased from Beyotime Institute of Biotechnology (Beyotime, China). Primary antibodies against heat shock protein (HSP) 90, HSP70, and HSP27 were purchased from Santa Cruz Biotechnology Inc. Primary antibody against β-actin and all secondary antibodies used for western blotting were purchased from Proteintech (Wuhan, China). Fluorescently tagged human monoclonal antibodies were purchased from BD Pharmingen (Becton Dickenson, San Jose, CA).

Metformin and Thioflavin T (ThT) reagent were purchased from Sigma-Aldrich (Saint Louis, MO, USA). A BCA protein assay kit and superoxide dismutase (SOD) assay kit were obtained from Beyotime Biotechnology. The malondialdehyde (MDA) Assay Kit (TBA method) was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). qPCR reagents and ECL kit were purchased from Invitrogen. All the antibodies were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA).