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MitoView™ Dyes

MitoView™ dyes are fluorogenic mitochondrial stains for live cells. The dyes rapidly accumulate in mitochondria and can be imaged without washing. They are available with blue, green, far-red, and near-infrared fluorescence.

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Product Description

MitoView™ dyes are fluorogenic mitochondrial stains for live cells. The dyes rapidly accumulate in mitochondria and can be imaged without washing. They are available with blue, green, far-red, and near-infrared fluorescence.

Features

  • Rapidly stain mitochondria in live cells without washing
  • Bright, photostable, and non-toxic
  • Monitor mitochondrial membrane potential with MitoView™ 633
  • Potential-independent MitoView™ Green can also stain fixed cells
  • MitoView™ 720 unique near-IR mitochondrial dye
  • Available in four colors

MitoView™ Dyes

ProductAbs/EmDetection channelPotential-dependent?Catalog no.Size
MitoView™ 405398/440 nmDAPIPartial†70070-T50 ug
7007020x50 ug
MitoView™ Green490/523 nmFITC, GFPNo70054-T50 ug
7005420x50 ug
MitoView™ 633622/648 nm*Cy®5, APC*Yes70055-T50 ug
7005520x50 ug
MitoView™ 720720/758 nm**Cy®5, Cy®7**Partial†70068-T50 ug
7006820x50 ug
* MitoView™ 633 also has red fluorescence in the Cy®3 channel and is not recommended for use with other red dyes.
**While optimal for Cy®7 settings, MitoView™ 720 is bright enough to be imaged in the Cy®5 channel.
† Dyes localize to the cytoplasm after mitochondrial depolarization, but still retain fluorescence.

Note: MitoView™ Blue (70052) has been discontinued and replaced by the improved MitoView™ 405. The original MitoView™ Blue will be available for purchase as a special order while supplies last. Contact techsupport@biotium.com to inquire about availability.

MitoView™ 405 is a blue fluorescent mitochondrial dye with absorbance/emission at 398/440 nm, suitable for detection by confocal microscopy or flow cytometry using settings for DAPI or Pacific Blue®. The dye is membrane permeable and becomes brightly fluorescent upon accumulation in the mitochondrial membrane. Mitochondrial localization is dependent on mitochondrial membrane potential; when membrane potential is disrupted the dye relocalizes to the cytoplasm, but still retains fluorescence. The dye is designed for use in live cells and is not fixable. MitoView™ 405 is a replacement for our original MitoView™ Blue, and has improved photostability.

MitoView™ Green is a green fluorescent mitochondrial dye with properties similar to those of MitoTracker® Green FM. The dye is non-fluorescent until it partitions into the mitochondrial membrane. The staining relies on mitochondrial mass, not on mitochondria membrane potential. Thus, the dye can be used to stain mitochondria in both live cells and fixed cells. MitoView™ Green is spectrally similar to FITC. Note: MitoView™ Green may be somewhat membrane potential-dependent in yeast cells.

MitoView™ 633 is a far-red fluorescent mitochondrial dye with absorbance/emission at 622/648 nm. The dye is membrane permeable and becomes brightly fluorescent upon accumulation in the mitochondrial membrane. Staining is dependent on mitochondrial membrane potential, and can be used to monitor mitochondrial membrane potential in intact cells. The dye is designed for use in live cells, and is not fixable. Note: The optimal detection settings for MitoView™ 633 are the same as for Cy®5 and other far-red dyes. However, the dye also has visible red fluorescence and can be imaged using Cy®3 settings as well. As a consequence, the dye cannot be used for two-color imaging with other red probes.

MitoView™ 720 is a near-infrared mitochondrial dye with absorbance/emission at 720/758 nm. While optimally detected using Cy®7 settings, the dye is bright enough to be imaged in the Cy®5 channel, and can be combined with visible red fluorescent probes. Mitochondrial localization is dependent on mitochondrial membrane potential; when membrane potential is disrupted the dye relocalizes to the cytoplasm but still retains fluorescence.

In addition to MitoView™ dyes, Biotium also offers several classical dyes for measuring mitochondrial membrane potential. These include JC-1, a ratiometric dye that forms red aggregates in healthy cells, which are reduced to the green monomeric state when mitochondrial membrane potential is lost. We also offer TMRM and TMRE, two red fluorescent dyes that are used to quantitatively measure mitochondrial membrane potential using the Nernst equation.

To view our wide selection of other cellular stains, visit our Cellular Stains technology page or see our Cellular Stains Brochure.

Cy Dye is a registered trademark of GE Healthcare. MitoTracker is a registered trademark of Thermo Fisher Scientific.

References

Mitoview™ 633 (70055)
1. Cell Death Discovery (2017), doi:10.1038/cddiscovery.2016.71
2. Biochimie (2017), doi: 10.1016/j.biochi.2017.06.004
3. Cell Death and Disease (2017), doi:10.1038/cddis.2017.307
4. Dalton Trans., (2017), doi: 10.1039/C7DT03836D
5. ACS Applied Materials& Interfaces (2018), doi: 10.1021/acsami.8b07917
6. PNAS (2018), www.pnas.org/cgi/doi/10.1073/pnas.1800526115
7. Chem (2018), https://doi.org/10.1016/j.chempr.2018.08.002

Mitoview™ Green (70054)
1. Analytical Chemistry (2018), doi: 10.1021/acs.analchem.8b03035
2. Cell Metabolism (2016), http://dx.doi.org/10.1016/j.cmet.2016.12.021
3. Human Molecular Genetics (2017), https://doi.org/10.1093/hmg/ddx319

 

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Support & Faq

Product shelf life, solubility, shipping and storage, and stability

Bioscience kits
The guaranteed shelf life from date of receipt for bioscience kits is listed on the product information sheet. Some kits have an expiration date printed on the kit box label, this is the guaranteed shelf life date calculated from the day that the product shipped from our facility. Kits often are functional for significantly longer than the guaranteed shelf life. If you have an older kit in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the kit still works for your application before processing a large number of samples or precious samples.

Antibodies and other conjugates
The guaranteed shelf life from date of receipt for antibodies and conjugates is listed on the product information sheet. Antibodies and other conjugates often are functional for significantly longer than the guaranteed shelf life. If you have an older conjugate in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples.

For lyophilized antibodies, we recommend reconstituting the antibody with glycerol and antimicrobial preservative like sodium azide for the longest shelf life (note that sodium azide is not compatible with HRP-conjugates).

Chemicals, dyes, and gel stains
Biotium guarantees the stability of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label. Fluorescent compounds should be protected from light for long term storage.

If you have a Biotium compound that has been in storage for longer than one year that you wish to use, we recommend performing a small scale positive control experiment to confirm that the compound still works for your application before processing a large number of samples or precious samples.

Expiration date based on date of manufacture (DOM)
If your institution requires you to document expiration date based on date of manufacture for reagents, please contact techsupport@biotium.com for assistance.

Chemical products with special stability considerations:

Esters

Ester compounds include the following:
• Succinimidyl esters (SE, also known as NHS esters), such as our amine-reactive dyes
• Acetoxymethyl esters (AM esters) such as our membrane-permeable ion indicator dyes
• Diacetate-modified dyes, like ViaFluor™ 405, CFDA, and CFDA-SE cell viability/cell proliferation dyes

Ester dyes are stable in solid form as long as they are protected from light and moisture. Esters are not stable in aqueous solution. Concentrated stock solutions should be prepared in anhydrous DMSO (see Biotium catalog no. 90082). Stock solutions in anhydrous DMSO can be stored desiccated at -20°C for one month or longer. Esters should be diluted in aqueous solution immediately before use. Succinimidyl esters (SE) should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound.

A note on CF™ dye succinimidyl ester stability
Succinimidyl esters are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Heavily sulfonated dyes, such as the Alexa Fluor® dyes, DyLight® dyes and IRDyes® are particularly hygroscopic, worsening the hydrolysis problem. For example, the percent of active Alexa Fluor® 488 succinimidyl ester (SE) could be well below 50% by the time of application (according to the manufacturer’s product datasheet). In a number of Alexa Fluor® SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while in all of Biotium’s CF™ dyes the SE group is prepared from an aliphatic carboxylic acid. This structural difference reduces the susceptibility of CF™ dye SE reactive groups to hydrolysis, resulting in relatively stable reactive dyes with consistently higher labeling efficiency compared to other SE derivatives of other fluorescent dyes.

Maleimides, MTS and thiosulfate dyes
Like the succinimidyl ester dyes, these dyes are also susceptible to hydrolysis, although generally to a much lower degree. Thus, for long term storage, anhydrous DMSO is recommended for making stock solutions.

Other reactive dyes
Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dye’s functional group. See the product information sheets for specific reactive dyes more information.

Coelenterazines and D-luciferin

Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also can be used as a solvent to minimize evaporation. If the solvent evaporates, the coelenterazine will still be present in the vial, so note the volume in the vial prior to storage so that you can adjust the solvent volume to correct for evaporation if needed. Prepare working solutions in aqueous buffers immediately before use. Coelenterazines are stable for up to five hours in aqueous solution.

Aquaphile™ coelenterazines are water soluble formulations of coelenterazines. They are stable in solid form when stored as recommended. Aquaphile™ coelenterazines should be dissolved in aqueous solution immediately before use. They are stable for up to five hours in aqueous solution.

Note that coelenterazines are predominantly yellow solids, but may contain dark red or brown flecks. This does not affect product stability or performance. If your coelenterazine is uniformly brown, then it is oxidized and needs to be replaced.

D-luciferin is stable in solid form and as a concentrated stock solution when stored as recommended; it is not stable at dilute working concentrations in aqueous solution. Prepare concentrated D-luciferin stock solutions (typically 1-100 mg/mL) in water, and store in aliquots at -20°C or below for six months or longer. Prepare working solutions immediately before use.

Category: Mix-n-Stain™ Antibody Labeling Kits, DNA Quantitation Kits, NucView® Caspase 3 Enzyme Substrates, Luciferase Assays, CF® Dyes, Product shelf life, solubility, shipping and storage, and stability

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Many of our solid compounds are packaged by lyophilization, in which case they usually do not appear as fluffy powders, but form a film or coating on the sides of the vial. Simply add the appropriate volume of the recommended solvent to the vial to make the desired concentration stock solution, and swirl or gently vortex to mix. Make sure the solvent comes in contact with the inside walls of the vial to fully recover the product.

Category: Product shelf life, solubility, shipping and storage, and stability

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Most of our products are stable at room temperature for many days, but we recommend storage at 4°C or -20°C to prolong shelf life. In the case of many of our aqueous dye solutions, the compounds are very stable at room temperature, but we recommend cold storage to prevent the growth of mold or other microbes over time. Therefore, to save on shipping costs, products with recommended storage at 4°C or -20°C may ship at ambient temperature or with an ice pack. These products may thaw without affecting product performance. When you receive the product, place it under the recommended storage conditions.

Category: Mix-n-Stain™ Antibody Labeling Kits, DNA Quantitation Kits, NucView® Caspase 3 Enzyme Substrates, Luciferase Assays, CF® Dyes, Product shelf life, solubility, shipping and storage, and stability

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Shipping/Shelf-life

Shipment Method: Shipping and handling methods will be assessed and calculated at time of shipment based upon item(s) storage temperature conditions. Please request expedited shipping in the Order Notes section if desired. Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.