2.2. Superoxide Dismutase (SOD) Assay Procedure

Muscle tissue perfusion was conducted in PBS buffer, at pH 7.4. Homogenization was executed in 5 mL of a 20 mM HEPES buffer (pH 7.2, 1 mM EDTA 210 mM mannitol, 70 mM sucrose per 1 g of tissue), chilled to 4 °C. After that, obtained homogenates were centrifuged at 2500× g for 15 min at the temperature of 4 °C. To prevent uncontrolled reaction initiation, it is crucial to store samples on ice until the analysis will be started. Assay procedure was performed with the Superoxide Dismutase Assay Kit, Item No. 706002 (Cayman Chemical Company; Ann Arbor, MI, USA). Measurement of absorbance was made with the help of a microplate reader Synergy4 (Biotek; Winooski, VT 05404 USA). Total activity of SOD was expressed in U/mL.