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MitoLite™ Green FM

Fluorescence images of HeLa cells stained with MitoLite™ Green FM using fluorescence microscope with a FITC filter set (Green). Live cells were co-stained with lysosome dye LysoBrite™ Red (Cat#22645, Red) and nuclei stain Nuclear Violet™ LCS1 (Cat#17543, Blue). 
Fluorescence images of HeLa cells stained with MitoLite™ Green FM using fluorescence microscope with a FITC filter set (Green). Live cells were co-stained with lysosome dye LysoBrite™ Red (Cat#22645, Red) and nuclei stain Nuclear Violet™ LCS1 (Cat#17543, Blue). 
Ordering information
Price ()
Catalog Number22695
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 weight671.88
SolventDMSO
Spectral properties
Excitation (nm)508
Emission (nm)528
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
201860-17-5
Molecular weight
671.88
Excitation (nm)
508
Emission (nm)
528
MitoLite™ Green FM is the same molecule to the MitoTracker Green FM (M7514, ThermoFisher). It is green-fluorescent mitochondrial stain. Unlike other MitoLite probes, MitoLite™ Green FM appears to localize to mitochondria, much less depending on mitochondrial membrane potential. The dye stains live cells, but it is not well-retained after aldehyde fixation.

Platform


Flow cytometer

Excitation488 nm laser
Emission530/30 nm filter
Instrument specification(s)FITC channel

Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall/clear bottom

Example protocol


AT A GLANCE

Protocol Summary

  1. Prepare 1 mM MitoLite™ Green FM stock solution
  2. Prepare 20-200 nM MitoLite™ Green FM staining solution
  3. Remove the growth media from the cells
  4. Add MitoLite™ Green FM staining solution to cells
  5. Incubate at 37°C for 30 minutes
  6. Wash cells and replace with 1x Hanks and 20mM Hepes Buffer (HH buffer)
  7. Observe cells using a fluorescence microscope with FITC filter set

Important  Bring the dye at room temperature before starting the experiment.

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™ Green FM stock solution:
Dissolve one vial MitoLite™ Green FM (50 ug) in 74 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™ Green FM staining solution:
Dilute 1 mM MitoLite™ Green FM stock solution to the final working concentration in HH buffer. The working concentration can be in the range of 20–200 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™ Green FM 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 FITC 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™ Green FM 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 flow cytometry (530/30 nm filter- FITC channel) or fluorescence microscopy (FITC filter set).

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of MitoLite™ Green FM 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 mM148.836 µL744.181 µL1.488 mL7.442 mL14.884 mL
5 mM29.767 µL148.836 µL297.672 µL1.488 mL2.977 mL
10 mM14.884 µL74.418 µL148.836 µL744.181 µL1.488 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)508
Emission (nm)528

Citations


View all 1 citations: Citation Explorer
Sonodynamic Therapy Promotes Efferocytosis via CD47 Down-Regulation in Advanced Atherosclerotic Plaque
Authors: Cao, Yang and Yao, Jianting and Gao, Weiwei and Cao, Zhengyu and Diabakte, Kamal and Wang, Linxin and Sun, Xin and Tian, Ye
Journal: International Heart Journal (2022): 21--233

References


View all 5 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