Cat activity assay kit

SOD activity was evaluated by using SOD activity assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to manufacture’ s instruction, based on the auto-oxidation of hydroxylamine, with the developed blue color then measured at 560 nm. Results are expressed as units of SOD/mg protein and calculated based on the formula: SOD activity (U/mg) = (A_light −A_measure)/50% × A_light × V_tissue × tissue mass (mg).
CAT activity was detected by using CAT activity assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to manufacture’ s instruction. The decomposition of H₂O₂ by CAT was stopped by the addition of ammonium molybdate. The remaining H₂O₂ was then reacted with ammonium molybdate to generate a pale-yellow complex, which was measured at 405 nm (Wei et al., 2018 (link)). The CAT activity was calculated based on the formula: CAT activity (U/mg) = [(Aself-control − A measure)/Ablank] × 650/protein concentration(mg / ml). The protein concentration of each sample was determined using BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA).

Catalase (CAT), Malondialdehyde (MDA), Peroxidase (POD) and Superoxide dismutase (SOD) were measured using Nanjing Jiancheng Bioengineering Institute’s relevant kits following manufacturer’s instructions and well detailed by Tang et al. [83 ]. Peroxidase (POD, E.C. 1.11.1.7) activity was assayed by peroxidase assay kit (Catalog No.A084; Jiancheng Bioengineering Institute, Nanjing, China). POD can catalyze the reaction of hydrogen peroxide, the enzyme activity of POD was obtained by measuring the change of absorbance at 420 nm. Superoxide dismutase (SOD, E.C. 4 1.15.1.1) activity was measured using a superoxide dismutase activity assay kit (Catalog No. A001–1; Jiancheng Bioengineering Institute, Nanjing, China). SOD activity was determined by the xanthine oxidase method (hydroxylamine). Catalase (CAT, E.C. 1.15.1.1) activity was measured using a CAT activity assay kit (Catalog No. A007–1; Jiancheng Bioengineering Institute, Nanjing, China). CAT can decompose H₂O₂ and this reaction can be quickly suspended by adding ammonium molybdate, the rest of H₂O₂ combine with ammonium molybdate to produce a pale-yellow complex compound, which is detected at 405 nm. Malondialdehyde (MDA) was measured using the MDA assay kit (TBA method) (Catalog No. A003–1; Jiancheng Bioengineering Institute, Nanjing, China) The MDA content was measured at 532 nm in nmol mg^− 1 proteins.