Are MitoSOX Mitochondrial Superoxide Indicators fixable?
MitoSOX indicator-stained cells should be imaged within 2 hr and are not fixable.
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What is the shipping temperature for MitoSOX Mitochondrial Superoxide Indicators, for live-cell imaging?
MitoSOX Mitochondrial Superoxide Indicators, for live-cell imaging are shipped at ambient temperature and should be stored as recommended.
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Why was the excitation wavelength for MitoSOX Red (Cat. Nos. M36008, M36007, M36009) changed from 510 nm to 396 nm in the new manual? What is the optimal wavelength to measure the signal?
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.
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Can I use MitoSOX Mitochondrial Superoxide Indicators (Cat. Nos. M36005, M3006, M36007, M36008, M36009) with other ROS reagents that don't accumulate within the mitochondria?
We do not recommend using MitoSOX Mitochondrial Superoxide Indicators (Cat. Nos. M36005, M3006, M36007, M36008, M36009) with other ROS reagents. Other ROS reagents may compete for radicals in or around the mitochondria limiting the response of the MitoSOX reagents.
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Can I store stock solutions of MitoSOX Mitochondrial Superoxide Indicators (Cat. Nos. M36005, M3006, M36007, M36008, M36009) beyond one day?
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.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.