MitoLite™ Red CMXRos
MitoLite™ Red CMXRos is chemically same to the MitoTracker™ Red CMXRos (ThermoFisher). MitoLite™ Red CMXRos is a cationic dye that selectively accumulates in mitochondria probably vial the mitochondrial membrane potential gradient. The mitochondrial indicator is a hydrophobic compound that easily permeates intact live cells, and trapped in mitochondria after it gets into cells. This fluorescent mitochondrial indicator is retained in mitochondria for long time since the indicator carries a cell-retaining group. This key feature significantly increases its staining efficiency. MitoLite™ Red CMXRos is well-retained after aldehyde fixation.
Example protocol
AT A GLANCE
Protocol Summary
- Prepare 1 mM MitoLite™ Red CMXRos stock solution
- Prepare 100-500 nM MitoLite™ Red CMXRos staining solution
- Remove the growth media from the cells
- Add MitoLite™ Red CMXRos staining solution to cells
- Incubate at 37°C for 30 minutes
- Wash cells and repalce with 1x Hanks and 20mM Hepes Buffer (HH buffer)
- Observe cells using a fluorescence microscope with TRITC filter set
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
MitoLite™ Red CMXRos stock solution:
Dissolve one vial MitoLite™ Red CMXRos (50 ug) in 94 uL high-quality, anhydrous dimethylsulfoxide (DMSO) to make 1 mM stock solution. Note: Keep the stock solution frozen at ≤–15°C and protected from light.
PREPARATION OF WORKING SOLUTION
MitoLite™ Red CMXRos staining solution:
Dilute 1 mM MitoLite™ Red CMXRos stock solution to the final working concentration in HH buffer. Note: The working concentration can be in the range of 100–500 nM.
SAMPLE EXPERIMENTAL PROTOCOL
Staining adherent cells:
- Grow cells to reach the desired confluency.
- Remove the growth media from the cells.
- Add MitoLite™ Red CMXRos staining solution to each well.
- Incubate at 37°C for 30 minutes.
- Wash cells and replace with 1x Hanks and 20mM Hepes Buffer (HH buffer).
- Observe cells using a fluorescence microscope with TRITC filter set. Note: The staining protocols is good for HeLa cell line and it may need to be optimized with the particular cell types.
Staining suspension cells:
- Centrifuge cells to a pellet and aspirate the supernatant.
- Resuspend the cells gently in MitoLite™ Red CMXRos staining solution.
- Incubate at 37°C for 30 minutes.
- Centrifuge the cells, remove supernatant and resuspend cells in fresh HH buffer.
- Cells may be analyzed by fluorescence microscopy (TRITC filter set).
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of MitoLite™ Red CMXRos to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 188.14 µL | 940.698 µL | 1.881 mL | 9.407 mL | 18.814 mL |
5 mM | 37.628 µL | 188.14 µL | 376.279 µL | 1.881 mL | 3.763 mL |
10 mM | 18.814 µL | 94.07 µL | 188.14 µL | 940.698 µL | 1.881 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Open in Advanced Spectrum Viewer
Product family
Name | Excitation (nm) | Emission (nm) |
MitoLite™ Red FX600 | 580 | 598 |
Citations
View all 4 citations: Citation Explorer
Tetramerization of PKM2 alleviates traumatic brain injury by ameliorating mitochondrial damage in microglia
Authors: Zhu, Haiyan and Zhang, Huiwen and Zhao, Xiao-Jing and Zhang, Lingyuan and Liu, Xue and Zhang, Zhi-Yuan and Ren, Yi-Zhi and Feng, Yong
Journal: (2023)
Authors: Zhu, Haiyan and Zhang, Huiwen and Zhao, Xiao-Jing and Zhang, Lingyuan and Liu, Xue and Zhang, Zhi-Yuan and Ren, Yi-Zhi and Feng, Yong
Journal: (2023)
Chitosan Oligosaccharides Alleviate H2O2-stimulated Granulosa Cell Damage via HIF-1$\alpha$ Signaling Pathway
Authors: Yang, Ziwei and Hong, Wenli and Zheng, K and Feng, Jingyuan and Hu, Chuan and Tan, Jun and Zhong, Zhisheng and Zheng, Yuehui
Journal: Oxidative Medicine and Cellular Longevity (2022)
Authors: Yang, Ziwei and Hong, Wenli and Zheng, K and Feng, Jingyuan and Hu, Chuan and Tan, Jun and Zhong, Zhisheng and Zheng, Yuehui
Journal: Oxidative Medicine and Cellular Longevity (2022)
Flow cytometric readout based on Mitotracker Red CMXRos staining of live asexual blood stage malarial parasites reliably assesses antibody dependent cellular inhibition
Authors: Jogd, undefined and , P. S., Singh, S. K., Christiansen, M., Dziegiel, M. H., Singh, S., Theisen, M.
Journal: Malar J (2012): 235
Authors: Jogd, undefined and , P. S., Singh, S. K., Christiansen, M., Dziegiel, M. H., Singh, S., Theisen, M.
Journal: Malar J (2012): 235
Compartment-specific Ca2+ imaging in the green alga Chlamydomonas reinhardtii reveals high light-induced chloroplast Ca2+ signatures
Authors: Pivato, Matteo and Grenzi, Matteo and Costa, Alex and Ballottari, Matteo
Journal: New Phytologist
Authors: Pivato, Matteo and Grenzi, Matteo and Costa, Alex and Ballottari, Matteo
Journal: New Phytologist
References
View all 6 references: Citation Explorer
the influence of platelets and a comparison of analytical approaches
Authors: Hopkinson, K.; Williams, E. A.; Fairburn, B.; Forster, S.; Flower, D. J.; Saxton, J. M.; Pockley, A. G., A MitoTracker Green-based flow cytometric assay for natural killer cell activity: variability
Journal: Exp Hematol (2007): 350-7
Authors: Hopkinson, K.; Williams, E. A.; Fairburn, B.; Forster, S.; Flower, D. J.; Saxton, J. M.; Pockley, A. G., A MitoTracker Green-based flow cytometric assay for natural killer cell activity: variability
Journal: Exp Hematol (2007): 350-7
Efficacy of MitoTracker Green and CMXrosamine to measure changes in mitochondrial membrane potentials in living cells and tissues
Authors: Pendergrass, W.; Wolf, N.; Poot, M.
Journal: Cytometry A (2004): 162-9
Authors: Pendergrass, W.; Wolf, N.; Poot, M.
Journal: Cytometry A (2004): 162-9
Mitotracker green is a P-glycoprotein substrate
Authors: Marques-Santos, L. F.; Oliveira, J. G.; Maia, R. C.; Rumjanek, V. M.
Journal: Biosci Rep (2003): 199-212
Authors: Marques-Santos, L. F.; Oliveira, J. G.; Maia, R. C.; Rumjanek, V. M.
Journal: Biosci Rep (2003): 199-212
MitoTracker Green labeling of mitochondrial proteins and their subsequent analysis by capillary electrophoresis with laser-induced fluorescence detection
Authors: Presley, A. D.; Fuller, K. M.; Arriaga, E. A.
Journal: J Chromatogr B Analyt Technol Biomed Life Sci (2003): 141-50
Authors: Presley, A. D.; Fuller, K. M.; Arriaga, E. A.
Journal: J Chromatogr B Analyt Technol Biomed Life Sci (2003): 141-50
and MitoTracker Green is affected by mitochondrial membrane potential altering drugs
Authors: Keij, J. F.; Bell-Prince, C.; Steinkamp, J. A., Staining of mitochondrial membranes with 10-nonyl acridine orange, MitoFluor Green
Journal: Cytometry (2000): 203-10
Authors: Keij, J. F.; Bell-Prince, C.; Steinkamp, J. A., Staining of mitochondrial membranes with 10-nonyl acridine orange, MitoFluor Green
Journal: Cytometry (2000): 203-10
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