Cellular ROS Assay Kit (Orange) ab186028 uses an ROS sensor to quantify ROS in live cells.
Suspension cells, Adherent cells
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Cellular ROS Assay Kit (Orange) ab186028 uses an ROS sensor to quantify ROS in live cells.
Suspension cells, Adherent cells
Microplate reader, Fluor. microscope, Flow cyt.
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Cellular ROS Assay Kit (Orange) ab186028 uses an ROS sensor to quantify ROS in live cells. The orange dye used in the ROS assay protocol is cell-permeable and generates orange fluorescence when it reacts with ROS. The fluorescence signal of the dye can be measured by fluorescence microscopy, high-content imaging, microplate fluorometry, or flow cytometry.
ab186028 provides a sensitive fluorometric, one-step assay to detect intracellular ROS (especially superoxide and hydroxyl radical) in live cells within 1 hour. The ROS assay can be performed in a convenient 96-well or 384-well microtiter-plate format using either a fluorescence microplate reader at Ex/Em = 540/570 nm or a fluorescent microscope with TRITC filter.
Previously called Cellular Reactive Oxygen Species Detection Assay Kit (Orange Fluorescence).Reactive oxygen species (ROS) are natural byproducts of the normal metabolism of oxygen and play important roles in cell signaling. However, during oxidative stress-related states, ROS levels can increase dramatically. The accumulation of ROS results in significant damage to cell structures. The role of oxidative stress in cardiovascular disease, diabetes, osteoporosis, stroke, inflammatory diseases, a number of neurodegenerative diseases and cancer has been well established. The ROS measurement will help to determine how oxidative stress modulates varied intracellular pathways.**Related products**Review the , or the full to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.To measure reactive oxygen species within cells, we recommend . Alternative ROS assays are available in orange (ab186028), red (Cellular ROS Assay Kit (Red) ab186027), and deep red (Cellular ROS Assay Kit (Deep Red) ab186029). DCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates) ab238535 is used to measure ROS in biofluids, culture supernatants and cell lysates. For assays designed to differentiate ROS, superoxides, and reactive nitrogen species: to assay ROS and superoxides use ROS/Superoxide Detection Assay Kit (Cell-based) ab139476; to assay ROS, superoxides, and reactive nitrogen species use Cellular ROS/RNS Assay Kit ab139473; to assay superoxides use Mitochondrial Superoxide Assay Kit (Fluorometric) ab219943.
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Detection of ROS in Jurkat cells.
Jurkat cells were treated without (Blue) or with 100μM TBHP (Red) for 30min at 37 °C, and then loaded with ROS orange in a 5% CO2, 37 °C incubator for 1 hour. The fluorescent intensities were measured with a flow cytometer using FL2 channel
Detection of ROS in Hela cells.
Hela cells were seeded overnight at 15,000 cells/90 μl/well in a black wall/clear bottom 96-well plate. The cells were untreated (control) or treated with1 mM H2O2 or 100 μM TBHP for 30 minutes at 37 °C. The ROS Orange assay solution (100 μl/well) was added and incubated in a 5% CO2, 37°C incubator for 1 hour. The fluorescence signal was monitored at Ex/Em = 540/570 nm (cut off = 550 nm) with bottom read mode.
Hela cells stained with the Cellular Reactive Oxygen Species Detection Assay Kit (Orange Fluorescence) (ab186028) in a black wall/clear bottom 96-well plate. A: Untreated control cells. B: Cells treated with 100 μM TBHP for 30min before staining.
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