Superoxide dismutase sod activity assay kit

SLT extracts were provided in-kind by Shineway Pharmaceutical Group (Shijiazhuang, China). Hydrogen peroxide (H₂O₂), dimethylsulfoxide (DMSO), and trypan blue were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium Ham’s F-12 (DMEM/F12) (1:1 Mix) with L-Glutamine was purchased from Lonza (Basel, Switzerland). Fetal bovine serum (FBS) and penicillin and streptomycin (PS) were purchased from Gibco Life Technologies (Waltham, MA, USA). 3-(4,5-di-methylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) was purchased from Astral Scientific (Caringbah, Australia). The superoxide dismutase (SOD) activity assay kit was purchased from Cayman Chemical (Ann Arbor, MI, USA). CytoTox non-radioactive cytotoxicity assay was obtained from Promega Corporation (Madison, WI, USA). The cellular ROS/Superoxide detection assay kit was purchased from Abcam (Cambridge, UK). Anti-Bax, anti-Bcl-2, anti-cleaved caspase-3, and anti-beta actin antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). All other reagents and chemicals were of chemical analytical grade.

The antioxidant action of the iMSC secretome was measured by the Superoxide Dismutase (SOD) Activity Assay Kit (Cayman Chemical, Ann Arbor, MI, USA). Samples were assayed according to the manufacturer’s instructions. Absorbance was measured at 450 nm using a Varioskan Flash microplate reader (Thermo Electron, Vantaa, Finland). Units of SOD activity were calculated from a standard curve using purified bovine erythrocyte SOD enzyme. All measurements were performed in duplicate.