![]() The ROS-Glo™ Assay uses an H 2O 2 substrate that directly reacts with H 2O 2 to produce a luciferin precursor, which is not a suitable substrate for luciferase. Since various ROS are converted to H 2O 2 in the cell, and H 2O 2 is the longest-lived ROS, an increase in H 2O 2 can reflect a general increase in the ROS level. The ROS-Glo™ Assay also provides a simple format for both cell-based and enzymatic assays. Importantly, the ROS-Glo™ Assay does not use horseradish peroxidase (HRP), which is known to cause a high number of false hits. The ROS-Glo™ H 2O 2 Assay is a luminescent assay that detects H 2O 2 directly, minimizing the false hit rate. New assays are needed in order to study the biology of ROS and to screen chemical compounds for their capacity to alter H 2O 2 levels in cultured cells or in enzymatic reactions. Current cell-based ROS assays require large numbers of cells with sample manipulation and are not amenable to higher throughput applications. Ĭurrent fluorescent enzyme assay formats are prone to false hit rates that are too high for efficient screening applications. For example, superoxide dismutase rapidly converts superoxide to H 2O 2. H 2O 2 has a relatively long half-life in solution and can diffuse out of the cell, which makes it a good marker of oxidative stress. H 2O 2 is a biomarker of enzymatic activities that either consume or produce H 2O 2. Enzymatic and non-enzymatic reactions result in conversion of ROS species to hydrogen peroxide (H 2O 2) in cells ROS are reactive (e.g., superoxide, singlet oxygen, H 2O 2) and have a short half-life in solution. ROS can also lead to cellular damage, or oxidative stress, as a result of environmental factors (e.g., radiation) or aberrant metabolism ROS are beneficial to the cell, having roles in cell signaling and as natural byproducts of normal metabolism Reactive oxygen species (ROS) are chemically reactive molecules that contain oxygen. ![]()
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