About Elite™ Mitochondrial ROS Activity Assay Kit (Orange Fluorescence)

Reactive oxygen species (ROS) are chemically reactive oxygen-containing molecules generated primarily during mitochondrial oxidative phosphorylation. Under physiological conditions, mitochondrial ROS function as important signaling molecules that regulate cell proliferation, differentiation, immune responses, and cellular adaptation to stress. However, excessive production of ROS—particularly superoxide (O₂•⁻) and hydroxyl radicals (•OH)—causes oxidative damage to DNA, proteins, and lipids, ultimately leading to mitochondrial dysfunction, apoptosis, and numerous pathological conditions. Mitochondrial oxidative stress has been implicated in cancer, neurodegenerative disorders, cardiovascular disease, diabetes, aging, inflammation, and drug-induced toxicity, making accurate detection of mitochondrial ROS essential for biomedical research and drug discovery.

The Elite™ Mitochondrial ROS Activity Assay Kit (Orange Fluorescence) provides a rapid, sensitive, and homogeneous fluorometric assay for detecting mitochondrial ROS in live cells. The assay utilizes Elite™ ROS Orange, a cell-permeable, non-fluorescent probe that selectively accumulates within mitochondria and produces a bright orange fluorescent signal upon oxidation by superoxide and hydroxyl radicals. The simple mix-and-read workflow requires no wash steps and is compatible with fluorescence microplate readers (Ex/Em = 540/570 nm), fluorescence microscopy, and flow cytometry. Optimized for 96- and 384-well microplates, the kit enables robust, reproducible measurement of mitochondrial oxidative stress and is readily adaptable to high-throughput screening (HTS) applications.

Applications

Biological Significance of Mitochondrial ROS

Mitochondria are the primary intracellular source of reactive oxygen species, with superoxide generated as a natural byproduct of electron leakage from the mitochondrial electron transport chain. While physiological levels of ROS regulate numerous signaling pathways, excessive mitochondrial ROS production disrupts cellular redox homeostasis and contributes to oxidative damage, inflammation, apoptosis, and impaired energy metabolism. Elevated mitochondrial ROS have been linked to the progression of cancer, Alzheimer's disease, Parkinson's disease, cardiovascular disorders, diabetes, ischemia-reperfusion injury, and age-related diseases. Quantitative measurement of mitochondrial ROS therefore provides valuable insight into mitochondrial function, cellular stress responses, therapeutic mechanisms, and disease progression. The Elite™ Mitochondrial ROS Activity Assay Kit (Orange Fluorescence) offers a reliable, highly sensitive, and convenient platform for investigating mitochondrial oxidative stress in cell biology, pharmacology, toxicology, and biomedical research.

User's Guide for Elite™ ROS Activity Assay Kits

   *1-fluorescence microplate reader; 2- fluorescence microscope; 3- flow cytometry