The reduced glutathione levels were evaluated with the method illustrated by Siddiqui et al. (2021), while activities of the catalase (CAT), and glutathione-S-transferase (GST) were assayed using the methods of Toledo et al. (2011) (link). On the other hand, nitric oxide (NO) and the peroxidation marker of the lipids (MDA, malondialdehyde) were quantified following the referenced methods of Miranda et al. (2001) (link) and Fu et al. (2018) (link), respectively.
Moreover, the tumor necrosis factor-alpha (TNF-α) was estimated by Quantikine ELISA Immunoassay kit (R&D systems) as per the guidelines provided by the manufacturer. The main antibody was used to sensitize 96-well microplates for 30 min at room temperature. The tissue sample was then added, cultured for a further 30 min, and washed. After washing, a peroxidase-conjugated secondary antibody was applied and allowed to incubate. Then, an ELISA plate reader was used to calculate the cytokine concentration.
Moreover, the tumor necrosis factor-alpha (TNF-α) was estimated by Quantikine ELISA Immunoassay kit (R&D systems) as per the guidelines provided by the manufacturer. The main antibody was used to sensitize 96-well microplates for 30 min at room temperature. The tissue sample was then added, cultured for a further 30 min, and washed. After washing, a peroxidase-conjugated secondary antibody was applied and allowed to incubate. Then, an ELISA plate reader was used to calculate the cytokine concentration.
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