The OxiSelect™ ROS Assay Kit is a cell-based assay for measuring hydroxyl, peroxyl, and other reactive oxygen species activity within a cell. The assay employs the cell-permeable fluorogenic probe DCFH-DA, which diffuses into cells and is deacetylcated by cellular esterases into the non-fluorescent DCFH (Figure 1). In the presence of ROS, DCFH is rapidly oxidized to highly fluorescent DCF. Fluorescence is read on a standard fluorometric plate reader.
Assay Principle.
ROS in HeLa Cells Treated with Hydrogen Peroxide. 50,000 HeLa cells in a 96-well plate were pretreated with 1 mM DCFH-DA for 60 minutes at 37ºC. Cells were then treated with hydrogen peroxide for 20 minutes.