Superoxide dismutase (SOD) activity was determined using a superoxide dismutase (SOD) activity assay kit (BioVision, Abcam, Waltham, MA, USA) [60 (link)]. The kit utilizes a WST-1 molecule (Water-Soluble Tetrazolium 1) that generates a water-soluble formazan dye upon reduction by the superoxide anion, which is liberated following the addition of an enzyme solution present in the kit. The superoxide reduction rate is linearly correlated with the xanthine oxidase activity, and it is inhibited by SOD. SOD inhibition activity can be colorimetrically determined at 450 nm. Cells were treated with sera, and proteins were extracted and quantified as described in Section 2.6 . The supernatant of each sample and different blanks containing equal protein amounts were used to measure the SOD activity. Samples and blanks were read using a plate reader at 450 nm (GENios Plus microplate reader, Tecan, Männedorf, Switzerland). To calculate the SOD activity (as inhibition rate %), the following equation was used:
where ABlank1 is the absorbance of the solution without the sample, ABlank2 is the absorbance of the solution without the enzyme working solution, and ABlank3 is the absorbance of the solution without the enzyme working solution and the sample.
where ABlank1 is the absorbance of the solution without the sample, ABlank2 is the absorbance of the solution without the enzyme working solution, and ABlank3 is the absorbance of the solution without the enzyme working solution and the sample.
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