O2 content test kit

The content of ${\text{O}}_2^{-}$ and H₂O₂ was measured using ${\text{O}}_2^{-}$ Content Test Kit and H₂O₂ Assay Kit (Jian Cheng Bioengineering Institute, Nanjing, China), respectively; 0.5 g of sample was added with 4.5 ml physiological saline and ground into homogenate. After incubation on ice for 2 h, the mixture was centrifuged at 8,000 g for 30 min, and the supernatant was used. The corresponding reagent was added according to the manufacturer's instructions, and ${\text{O}}_2^{-}$ and H₂O₂ contents were determined by measuring the OD at 550 nm and 405 nm, respectively. Both ${\text{O}}_2^{-}$ and H₂O₂ contents were expressed as mmol/g Prot.

The content of O₂⁻ was measured by O₂⁻ Content Test Kit (No. A052, Jian Cheng Bioengineering Institute, Nanjing, China) following the protocol of manufacturer. The content of H₂O₂ was calculated by Tissue H₂O₂ assay Kit (No. A064, Jian Cheng Bioengineering Institute, Nanjing, China) according to Yang, Han, Han, Ackah, Li, Bi, Yang and Prusky [27 (link)]. Briefly, 0.5 g samples were homogenized in 4.5 mL normal saline to extract the H₂O₂, then the test was carried out following the protocol of manufacturer. The contents of O₂⁻ and H₂O₂ were expressed as mol/kg.