MitoLite™ Red CMXRos

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	531.52
Solvent	DMSO

Spectral properties

Excitation (nm)	578
Emission (nm)	598

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200

Overview SDS Protocol

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

Excitation	TRITC filter set
Emission	TRITC filter set
Recommended plate	Black wall/clear bottom

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

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Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
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Spectrum

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Spectral properties

Excitation (nm)	578
Emission (nm)	598

Product Family

Name	Excitation (nm)	Emission (nm)
MitoLite™ Red FX600	580	598

Images

Figure 1. Fluorescence images of HeLa cells stained with MitoLite™ Red CMXRos.
Hela cells were stained with 0.1uM MitoLite™ Red CMXRos in HH buffer at 37^oC 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).

Figure 2. Chemical structure for MitoLite™ Red CMXRos.

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)

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)

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

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

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