Product nameCellular ROS Assay Kit (Deep Red)
See all Oxidative Stress kits
Sample typeAdherent cells, Suspension cells
Assay time1h 00m
Cellular ROS Assay Kit (Deep Red) ab186029 uses an ROS sensor to quantify ROS in live cells. The ROS Deep Red dye is cell-permeable and generates Deep Red fluorescence when it reacts with ROS. The fluorescence signal of the ROS Deep Red Dye can be measured by fluorescence microscopy, high-content imaging, microplate fluorometry, or flow cytometry.
ab186029 provides a sensitive fluorometric, one-step assay to detect intracellular ROS (especially superoxide and hydroxyl radical) in live cells within 1 hour. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format using either a fluorescence microplate reader at Ex/Em = 650/675 nm or a fluorescent microscope with TRITC filter.
Previously called Cellular Reactive Oxygen Species Detection Assay Kit (Deep Red 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.
Review the oxidative stress marker and assay guide, or the full metabolism assay guide 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.
PlatformMicroplate reader, Fluor. microscope, Flow cyt.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 200 tests Assay Buffer 1 x 20ml DMSO 1 x 100µl ROS Deep Red Dye (Lyophilized) 1 vial
Jurkat cells were treated without (Blue) or with 100µM TBHP (Red) for 30min at 37 °C, and then loaded with ROS Deep Red in a 5% CO2, 37 °C incubator for 1 hour. The fluorescent intensities were measured with a flow cytometer using FL2 channel
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 Deep Red 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 = 650/675 nm (cut off = 665 nm) with bottom read mode.
Images of Hela cells stained with the Cellular Reactive Oxygen Species Detection Assay Kit (Deep Red Fluorescence) (ab186029) in a black wall/clear bottom 96-well plate. A: Untreated control cells. B: Cells treated with 100 µM TBHP for 30min before staining.
This product has been referenced in:
- Larasati YA et al. Curcumin targets multiple enzymes involved in the ROS metabolic pathway to suppress tumor cell growth. Sci Rep 8:2039 (2018). Read more (PubMed: 29391517) »
- Duffy CM et al. Microglial Immune Response to Low Concentrations of Combustion-Generated Nanoparticles: An In Vitro Model of Brain Health. Nanomaterials (Basel) 8:N/A (2018). Read more (PubMed: 29522448) »