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MitoLite™ Red CMXRos

Fluorescence images of HeLa cells stained with MitoLite™ Red CMXRos.<br> Hela cells were stained with 0.1uM MitoLite™ Red CMXRos in HH buffer at 37<sup>o</sup>C for 30mins, and then washed twice with HH bufer. Cells were imaged with fluorescence microscope using a TRITC filter set (Red). Nuclei were stain with Nuclear Violet™ LCS1 (Cat#17543) and viewed with DAPI filter set (Blue).
Fluorescence images of HeLa cells stained with MitoLite™ Red CMXRos.<br> Hela cells were stained with 0.1uM MitoLite™ Red CMXRos in HH buffer at 37<sup>o</sup>C for 30mins, and then washed twice with HH bufer. Cells were imaged with fluorescence microscope using a TRITC filter set (Red). Nuclei were stain with Nuclear Violet™ LCS1 (Cat#17543) and viewed with DAPI filter set (Blue).
Chemical structure for MitoLite™ Red CMXRos.
Ordering information
Price ()
Catalog Number22698
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
Molecular weight531.52
SolventDMSO
Spectral properties
Excitation (nm)578
Emission (nm)598
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


CAS
167095-09-2
Molecular weight
531.52
Excitation (nm)
578
Emission (nm)
598
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.

Platform


Fluorescence microscope

ExcitationTRITC filter set
EmissionTRITC filter set
Recommended plateBlack wall/clear bottom

Example protocol


AT A GLANCE

Protocol Summary

  1. Prepare 1 mM MitoLite™ Red CMXRos stock solution
  2. Prepare 100-500 nM MitoLite™ Red CMXRos staining solution
  3. Remove the growth media from the cells
  4. Add MitoLite™ Red CMXRos staining solution to cells
  5. Incubate at 37°C for 30 minutes
  6. Wash cells and repalce with 1x Hanks and 20mM Hepes Buffer (HH buffer)
  7. 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:

  1. Grow cells to reach the desired confluency.

  2. Remove the growth media from the cells.

  3. Add MitoLite™ Red CMXRos staining solution to each well.

  4. Incubate at 37°C for 30 minutes.

  5. Wash cells and replace with 1x Hanks and 20mM Hepes Buffer (HH buffer).

  6. 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:

  1. Centrifuge cells to a pellet and aspirate the supernatant.

  2. Resuspend the cells gently in MitoLite™ Red CMXRos staining solution.

  3. Incubate at 37°C for 30 minutes.

  4. Centrifuge the cells, remove supernatant and resuspend cells in fresh HH buffer.

  5. 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 mg0.5 mg1 mg5 mg10 mg
1 mM188.14 µL940.698 µL1.881 mL9.407 mL18.814 mL
5 mM37.628 µL188.14 µL376.279 µL1.881 mL3.763 mL
10 mM18.814 µL94.07 µL188.14 µL940.698 µL1.881 mL

Molarity calculator

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Spectrum


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spectrum

Spectral properties

Excitation (nm)578
Emission (nm)598

Product family


NameExcitation (nm)Emission (nm)
MitoLite™ Red FX600580598

Citations


View all 1 citations: Citation Explorer
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

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
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
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
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
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
Detection of apoptosis in live cells by MitoTracker red CMXRos and SYTO dye flow cytometry
Authors: Poot M, Gibson LL, Singer VL.
Journal: Cytometry (1997): 358