Reactive oxygen species assay kit uses a unique fluorescent probe H2DCFDA to detect reactive oxygen species. H2DCFDA is chemically reduced and acetylated form of 2′,7′-dichloro- fluorescein (DCF) and is nonfluorescent and cell-permeant.
Once H2DCFDA enters into cells, the acetate groups are removed by intracellular esterases to form H2DCF. H2DCF is well retained inside cells. Oxidation of H2DCF by reactive oxygen species yields fluorescent DCF. Reactive oxygen species can be detected by monitoring the increase in fluorescence with a flow cytometer, fluorometer, microplate reader, or fluorescence microscope, using excitation sources and filters appropriate for fluorescein (FITC). Because the dyes are susceptible to photo-oxidation, low light conditions should be used for fluorescence microscopy applications whenever possible.
Figure 1. Detection of ROS with ROS assay kit (Cat No. A057).
Mol Cancer Ther (2013) 12:2446-2458