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Invitrogen™ MitoSOX™ Mitochondrial Superoxide Indicators, for live-cell imaging
Description
MitoSOX Green and MitoSOX Red superoxide indicators are novel fluorogenic dyes specifically targeted to mitochondria in live cells. Oxidation of the MitoSOX reagent by mitochondrial superoxide produces bright green or red fluorescence.
Features of these indicators include:
• Readily oxidized by superoxide but not by other ROS- or RNS-generating systems
• Use for live cell imaging
• Rapidly and selectively targeted to the mitochondria
• MitoSOX Green indicator: absorption/emission maxima of ∼488/510 nm (traditional FITC/GFP filter set)
• MitoSOX Red indicator: absorption/emission maxima of ∼396/610 nm* (custom filter set recommended)
*Note that two excitation peaks may be observed for MitoSOX Red indicator, as it is excited at both 510 nm and 396 nm. While 510 nm will excite the superoxide oxidation product, it can also excite non-specific products, so we recommend using a 396-nm excitation for more selective detection of mito superoxide when using MitoSOX Red indicator (1).
MitoSOX Green indicator is offered as one vial or a pack of five vials. MitoSOX Red indicator is offered as one vial or a pack of ten vials. A variety pack of one vial each of MitoSOX Red and MitSOX Green indicators is also available.
Consult user Manual for solubility instructions.
Quick, easy detection of mitochondrial superoxide in live cells
The production of superoxide by mitochondria can be visualized in fluorescence microscopy using MitoSOX superoxide indicators. They permeate live cells where they selectively target mitochondria. They are rapidly oxidized by superoxide but not by other reactive oxygen species (ROS) and reactive nitrogen species (RNS). The oxidized product is highly fluorescent.
MitoSOX indicators may be used to distinguish artifacts of isolated mitochondrial preparations from direct measurements of superoxide generated in the mitochondria of live cells. They may also provide a valuable tool in the research of agents that modulate oxidative stress in various pathologies. In addition, these indicators have been used in metabolic flux assays using high-content instruments (2). MitoSOX indicators have also been used to detect mitochondrial superoxide using flow cytometery (3).
References
1: Robinson, Kristine M., et al. Selective fluorescent imaging of superoxide in vivo using ethidium-based probes. Proceedings of the National Academy of Sciences 103.41 (2006): 15038-15043.
2: Little, Andrew Charles, et al. High-content fluorescence imaging with the metabolic flux assay reveals insights into mitochondrial properties and functions. Communications biology 3.1 (2020): 1-10.
3: Kauffman, Megan E. et al. MitoSOX-Based Flow Cytometry for Detecting Mitochondrial ROS. Reactive oxygen species (Apex, N.C.) vol. 2,5 (2016): 361-370.
Order Info
Shipping Condition: Room Temperature
Specifications
Specifications
| Color | Red |
| Format | Vial(s) |
| Quantity | 10 vials x 50 μg |
| Detection Method | Fluorescence |
| Excitation/Emission | ∼396/610 nm |
| Indicator | MitoSox Red |
| Product Line | MitoSOX |
Frequently Asked Questions (FAQs)
MitoSOX indicator-stained cells should be imaged within 2 hr and are not fixable.
MitoSOX Mitochondrial Superoxide Indicators, for live-cell imaging are shipped at ambient temperature and should be stored as recommended.
The excitation wavelength for MitoSOX Red (Cat. Nos. M36008, M36007, M36009) was changed from 510 nm to 396 nm in the new manual because we have found that excitation at 396 nm selectively excites the superoxide oxidation product without exciting other non-specific products. Although the indicator displays an adequate signal when excited at 510 nm, we highly recommend using the 396 nm excitation wavelength for selective detection of the indicator’s response to mitochondrial superoxide. For more information on the selective detection of superoxide at 396 nm, please refer to the article linked below.
Robinson KM, Janes MS, Pehar M, Monette JS, Ross MF, Hagen TM, Murphy MP, Beckman JS. Selective fluorescent imaging of superoxide in vivo using ethidium-based probes. Proc Natl Acad Sci U S A. 2006 Oct 10;103(41):15038-43. doi: 10.1073/pnas.0601945103. Epub 2006 Oct 2. PMID: 17015830; PMCID: PMC1586181.
The optimal emission wavelength to observe the superoxide oxidation product is 580 nm. Please note that the 610 nm wavelength mentioned in the MitoSOX Green and MitoSOX Red Mitochondrial Superoxide Indicators User Guide is incorrect.
We suggest a working concentration range of 100 nM to 5 µM. It is recommended to optimize the final working concentration for each sample and application. It is important to be cautious when using concentrations >1 µM, as higher non-specific labeling may occur as well as possible mitochondrial toxicity.
No. We do not recommend storing MitoSOX Mitochondrial Superoxide Indicators (Cat. Nos. M36005, M3006, M36007, M36008, M36009) stock solutions beyond one day as they are more susceptible to spontaneous oxidation (from radicals in the atmosphere) when in solution compared to storing the reagents in solid form.
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