I have been performing protein oxidation assay (using Oxyblot from Millipore) for a couple of weeks now. I irradiate my cells (lung cells) with a high dose of radiation, after which I extract the proteins (whole cell lysate) and perform this assay. However, I always see very high signal even in my untreated cells. I was wondering if anyone has performed this assay? If so, do you get the same high amount of oxidized proteins in the untreated cells (since all cells have basal level of oxidation)? I have looked at other articles and I found that most of them reported only small amount of oxidized proteins in the untreated sample. Thanks-
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