Hallo everyone,
I am trying to decrease ROS by adding glutathione (reduced) in human retinal endothelial cells under hyperoxic conditions. Unfortunately a lot of cellular ros assay kits like H2DCFDA are based on fluorogenic dyes which react with glutathione. Does anyone have experience or an idea how to measure oxidative stress and/or ROS based on reactions without interference with strong ROS scavenger like glutathione?
Thank you in advance
Justus
Justus S. Ernesti came across this paper for your question, though not the accurate one but to give you an idea
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6434272/
srinivas kasulla
I have done extensive analysis of oxidative stress in cells, but I found at that time there is no way to distinguish species of ROS. There are assessments of bulk ROS and measuring the damage directly. Depends on what you want to see, measuring the oxidative damage would be more directly address your question in general. There used to be a very good introduction as to which markers could visualize ROS. If you want to find ROS in mitochondria or elsewhere, there should be markers. Search the web to see if they still have those. I have not drafted human cells but here are some mouse and C. elegans papers but you might be interested:
Article Stress resistance in long-lived mouse models
Article Current Advances in the Studies of Oxidative Stress and Age-...
Article Multiplex stress resistance in cells from long-lived dwarf mice
By calculating the amount reactive species before and after use of glutathione in sample
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