Description
• Quick ~1 hour protocol
• Highly sensitive to 10 pM
• Detects the presence of various ROS species
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.
Introduction: Accumulation of reactive oxygen species (ROS) coupled with an increase in oxidative stress has been implicated in the pathogenesis of several disease states. The role of oxidative stress in vascular diseases, diabetes, renal ischemia, atherosclerosis, pulmonary pathological states, inflammatory diseases, and cancer has been well established. Free radicals and other reactive species are constantly generated in vivo and cause oxidative damage to biomolecules, a process held in check by the existence of multiple antioxidant and repair systems as well as the replacement of damaged nucleic acids, proteins and lipids. Measuring the effect of antioxidant therapies and ROS activity intracellularly is crucial to suppressing or treating oxidative stress inducers.
MyBioSource's OxiSelect Intracellular ROS Assay Kit (Green Fluorescence) is a cell-based assay for measuring hydroxyl, peroxyl, or other reactive oxygen species activity within a cell. The assay employs the cell-permeable fluorogenic probe 2', 7'-Dichlorodihydrofluorescin diacetate (DCFH-DA). In brief, DCFH-DA is diffused into cells and is deacetylated by cellular esterases to non-fluorescent 2', 7'-Dichlorodihydrofluorescin (DCFH), which is rapidly oxidized to highly fluorescent 2', 7'-Dichlorodihydrofluorescein (DCF) by ROS (Figure 1). The fluorescence intensity is proportional to the ROS levels within the cell cytosol. The effect of antioxidant or free radical compounds on DCF-DA can be measured against the fluorescence of the provided DCF standard. The kit has a DCF detection sensitivity limit of 10 pM. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples