Good morning, I am Miguel Alfaro.
As part of my thesis project, I am evaluating the antioxidant activity of the glutathione peroxidase enzyme (GPX) obtained from hemolyzed human erythrocytes, previously treated with different concentrations of a plant extract. For this assay, I am using the Glutathione Peroxidase Assay Kit from Cayman Chemicals (USA) (Item No. 703102) and following manufacturer´s instructions. However, when I performed these experiments, it is not detected any GPX enzyme activity in the experimental groups, including the negative control where it is supposed to find GPX activity because there is not any influence of a treatment. Only this activity in detected in the positive control (GPX bovine enzyme included in the kit).
It is important to mention that these samples of supernatants from erythrocytes were previously used for the evaluation of antioxidant activity of superoxide dismutase and catalase enzymes, using commercial kits from Cayman Chemicals and in which the antioxidant activity of these two enzymes were detected in all experimental groups.
For this reason, I would like to ask if someone has worked previously with this type of assay employing the same or a similar kit and that could give me some recommendations for my experiments.
I will be very grateful for your help. Thanks for your attention.
Check https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2657797/
Also check https://journals.sagepub.com/doi/pdf/10.1258/000456303766477002
See this article
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