Mitochondrial Superoxide Detection Kit (Fluorometric) (ab219943) is a sensitive fluorometric one-step assay to detect intracellular superoxide radical in live cells.
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- Frontiers in oncology 12:9038742022AXL Promotes Metformin-Induced Apoptosis Through Mediation of Autophagy by Activating ROS-AMPK-ULK1 Signaling in Human Esophageal Adenocarcinoma.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJun Hong,Selma Maacha,Nataliya Pidkovka,Andreia Bates,Safia N Salaria,Mary K Washington,Abbes BelkhiriPubMed 35936716
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Key facts
- Detection method
Fluorescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Cell-based
- Reactive species
Mammals
What's included?
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Mitochondrial Superoxide Detection Kit (Fluorometric) (ab219943) is a sensitive fluorometric one-step assay to detect intracellular superoxide radical in live cells.
Key facts
- Detection method
Fluorescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Cell-based
- Reactive species
Mammals
- Assay Platform
Microplate reader, Fluorescence microscope
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
- Storage information
-20°C
Notes
Mitochondrial Superoxide Detection Kit (Fluorometric) (ab219943) is a sensitive fluorometric one-step assay to detect intracellular superoxide radical in live cells. The assay uses our MitoROS 580 dye: the dye is cell-permeable and selectively reacts with mitochondrial superoxide present in live cells to generate a red fluorescence signal that can be easily read at Ex/Em = 540/590 nm.
The assay can be performed within one hour and can be detected by fluorescence microscopy, microplate reader or high-content imaging. It can be easily adapted to use in 384-well microplate format.
Mitochondria are major producers of cellular superoxide. The production of low to moderate levels of superoxide is critical for the proper regulation of many essential cellular processes including gene expression, signal transduction, and muscle adaptation to endurance exercise training. Uncontrolled mitochondrial superoxide production can trigger cellular oxidative damage that contributes to the pathogenesis of a wide variety of disorders including cancer, cardiovascular diseases, neurodegenerative diseases and aging. The detection of intracellular mitochondrial superoxide is of central importance to understanding proper cellular redox regulation and the impact of its dysregulation on various pathologies.**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 (Cellular ROS Assay Kit (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 ab219943.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Mitochondrial superoxide also known as mitosox refers to the reactive oxygen species (ROS) generated specifically in mitochondria. Superoxide a byproduct of the mitochondrial electron transport chain is a small molecule with a molecular mass of 31.9988 g/mol. The compound primarily emerges during cellular respiration in the mitochondria's inner membrane. Mitochondrial superoxide's excessive production can result in cellular damage since it acts as a precursor to more harmful ROS like hydrogen peroxide and hydroxyl radical.
- Biological function summary
Mitochondrial superoxide contributes to a cell’s oxidative state regulation. It plays an important role in various signaling pathways and in maintaining cellular homeostasis. Mitochondrial superoxide is often part of larger complexes involved in ROS detection such as the mitochondria itself that regulate ROS levels to ensure balance. These complexes can trigger signaling cascades that activate reparative or protective cellular responses making mitochondrial ROS pivotal players in maintaining cellular integrity.
- Pathways
Mitochondrial superoxide influences important metabolic processes such as the antioxidant defense and apoptosis pathways. ROS detection by mitochondria can modulate the balance between cell survival and cell death by interacting with proteins like cytochrome c and members of the Bcl-2 family. These proteins help determine whether a cell will undergo apoptosis in response to mitochondrial stress signals contributing essential regulation in oxidative stress pathways.
- Associated diseases and disorders
Mitochondrial superoxide holds a significant connection to neurodegenerative diseases and cardiovascular disorders. Excessive mitochondrial ROS is implicated in the oxidative stress that characterizes conditions like Parkinson’s disease and heart failure. In these contexts the interaction of mitochondrial superoxide with proteins such as SOD2 (superoxide dismutase 2) and NOX (NADPH oxidases) is critical. These proteins can modulate the damaging effects of superoxide and offer potential therapeutic targets for conditions linked to mitochondrial dysfunction.
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2 product images
Fluorescence Microscopy - Mitochondrial Superoxide Assay Kit (Fluorometric) (ab219943)
Superoxide production in HeLa cells. HeLa cells were seeded overnight (105 cells/well/100 μL) in a 96 well black wall/clear bottom plate. Left: cells were treated with 50 μM Antimycin A (AMA) at 37°C for 30 minutes, then incubated with MitoROS 580 for 1 hour. Right: control HeLa cells were incubated with MitoROS580 at 37 °C for 1 hour without treatment. The fluorescence signal was measured using fluorescence microscope with a TRITC filter.
Functional Studies - Mitochondrial Superoxide Assay Kit (Fluorometric) (ab219943)
Quantification of superoxide production in HeLa cells. HeLa cells were seeded overnight (105 cells/well/100 μL) in a 96 well black wall/clear bottom plate. Cells were left untreated (control) or treated with either pyocyanin (Pyo, 50 μM Pyocyanin) or antimycin A (AMA, 50 μM Antimycin A) at 37 °C for 30 minutes. Cells were then incubated with MitoROS 580 at 37 °C for 1 hour. The fluorescence signal was monitored at Ex/Em = 540/590 nm (cut off = 570 nm) with bottom read mode using a microplate reader.
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