Mitochondrial Superoxide Assay Kit (Fluorometric)
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(12 Publications)
- FuncS
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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.
- Fluorescence Microscopy
Supplier Data
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.
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Product details
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.
Other Notes
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
To measure reactive oxygen species within cells, we recommend DCFDA assay kit ab113851
Alternative ROS assays are available in orange (ab186028), red (ab186027), and deep red (ab186029)
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 ab139476
- to assay ROS, superoxides, and reactive nitrogen species use ab139473
- to assay superoxides use ab219943
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Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
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Publications (12)
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Biomedicines 13: PubMed40426844
2025
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Cancers 17: PubMed40361395
2025
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PLoS pathogens 21:e1012561 PubMed40258014
2025
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Redox biology 77:103348 PubMed39332053
2024
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Journal of translational medicine 22:221 PubMed38429788
2024
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Journal of the American Heart Association 12:e029131 PubMed37345832
2023
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Frontiers in oncology 12:903874 PubMed35936716
2022
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Frontiers in cell and developmental biology 9:688607 PubMed34381775
2021
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Cell death discovery 6:110 PubMed33133645
2020
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EMBO molecular medicine 12:e12423 PubMed32875720
2020
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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