I am researching on techniques that can measure molecular interactions, besides isometric titration calorimetry (ITC), because I would like to check if an antioxidant of my interest is interacting with my oxidant (hydrogen peroxide) before entering the cell and, with that, "make up" the effect of this antioxidant. And one of these techniques that I found was the anisotropy, and in it is used a fluorophore, such as FITC. Can the peroxide react or affect the bond with the FITC and produce other compounds?
Fluorescein (which is the fluorescent part of FITC; isothiocyanate is the amine-reactive part) is easily bleached by photochemical reactions. I would expect this to be enhanced by reactive oxygen species. You can easily test this by making a dilute solution of fluorescein, adding hydrogen peroxide, and measuring the effect of H2O2 concentration of the rate of photobleaching using a spectrofluorometer.
This paper may shed some light on the subject:
https://www.sciencedirect.com/science/article/abs/pii/S101113441400267X
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