Glutathione peroxidase gpx kit

Protein concentrations were measured by a BCA assay kit (Pierce, Jiancheng Bioengineering Institute, Nanjing, China). Guaiacol oxidation by MPO-produced hydroxyl radicals in the presence of hydrogen peroxide was evaluated as previously described [33 (link)]. This activity, expressed in units per mg of protein, was defined as the quantity of enzyme degrading 1 μmol of hydrogen peroxide per minute. The colon sample sections were mechanically homogenized on ice in a 100 mM Trizma buffer pH 7.4 with 1 mM EDTA and 1 mM phenylmethanesulfonyl fluoride (PMSF). The homogenate was sonicated on ice for thirty seconds and subsequently centrifuged at 1,000 x g for ten minutes at 4°C. The supernatants were then aliquoted and stored at -80 °C until subsequent analysis. Antioxidant enzyme activities, defined as the quantity degrading 1 μmol per minute of substrate (in units per mg proteins), were ascertained in triplicate using catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GRed) and glutathione peroxidase (GPx) kits (Jiancheng Bioengineering Institute, Nanjing, China).

The total antioxidant capacity and antioxidant enzyme activity was conducted using the manual for T-AOC kit and Glutathione peroxidase (GPX) kit which were purchased from Nanjing Jiancheng Technology Co., Ltd. The catalase activity test was performed using Catalase (CAT) kit and the superoxide dismutase activity was conducted following Superoxide dismutase (SOD) kit purchased from Beijing Solarbio Technology Co., Ltd.