The assessment of mitochondrial respiratory chain (MRC) activities entailed the use of both the MRC Complex Activity Assay Kit (Solarbio) and the ATP Synthetase Activity Assay Kit (YBio), adhering to the guidelines provided by the manufacturer. MRC activities were appraised via a colorimetric assay at 340, 550, and 450 nm, specifically targeting MRC complex I, MRC complex III, and ATP synthetase, respectively. The unit enzyme activity was quantified in terms of nmol/min/mg protein.
The evaluation of catalase activity was conducted using a readily available assay kit (Solarbio) in accordance with the provided guidelines. The preparation of protein samples involved lysing, and the total lysate protein concentration was determined through a BCA protein assay kit (Beyotime). Subsequently, each lysate was mixed with a colorimetric assay substrate solution from the kit and incubated at room temperature for a duration of 10 min. The absorbance at 405 nm was subsequently measured using a microplate reader (Thermo, MK3) along with a standard curve for reference.
The evaluation of catalase activity was conducted using a readily available assay kit (Solarbio) in accordance with the provided guidelines. The preparation of protein samples involved lysing, and the total lysate protein concentration was determined through a BCA protein assay kit (Beyotime). Subsequently, each lysate was mixed with a colorimetric assay substrate solution from the kit and incubated at room temperature for a duration of 10 min. The absorbance at 405 nm was subsequently measured using a microplate reader (Thermo, MK3) along with a standard curve for reference.