What does MitoTracker red stain for?
MitoTracker Red CMXRos is a red-fluorescent dye that stains mitochondria in live cells and its accumulation is dependent upon membrane potential. The dye is well-retained after aldehyde fixation.
Can you fix MitoTracker?
For MitoTracker® Red CMXRos, we have found that fixing with 3.7% formaldehyde in complete growth medium at 37°C for 15 minutes works well for endothelial cells.
How long does MitoTracker last?
The experiment will last approximately 48 hours.
How do you use the red MitoTracker?
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- Remove medium from the cells and add prewarmed medium with MitoTracker (most commonly used concentration is in the range of 300-400 nmol).
- Incubate for 45 to 60 minutes.
- Exchange the medium with regular one and incubate for 5 min, repeat 2 more times.
How does a MitoTracker work?
MitoTracker is a commercially available fluorescent dye (Invitrogen/Molecular Probes) that, like the aforementioned dyes, labels mitochondria within live cells utilizing the mitochondrial membrane potential. The dye becomes permanently bound to the mitochondria, and thus remains after the cell dies or is fixed.
How does MitoTracker green work?
MitoTracker Green (MTG) is a mitochondrial-selective fluorescent label commonly used in confocal microscopy and flow cytometry. It is expected that this dye selectively accumulates in the mitochondrial matrix where it covalently binds to mitochondrial proteins by reacting with free thiol groups of cysteine residues.
How do you dissolve a MitoTracker?
Dissolve one vial (50 µg) of MitoTracker Green FM (Invitrogen M7514) in DMSO to prepare a stock solution of 1 mm. Cover with aluminum foil to minimize light exposure. Store at or below −20°C.
How does MitoTracker deep red work?
MitoTracker® Deep Red binds thiol‐reactive chloromethyl groups in the mitochondrial membrane and is retained after cell fixation, whereas MitoTracker® Green binds free thiol groups of cysteine residues belonging to mitochondrial proteins and can only be used with non‐fixed cells.
What does MitoTracker measure?
MitoTracker green (MTG) and nonylacridine orange (NAO) are fluorescent probes which have been commonly used to assess mitochondrial mass. This is based on the assumption that both MTG and NAO accumulate in mitochondria regardless of the mitochondrial transmembrane potential (ΔΨm).
What does MitoTracker green stain?
MitoTracker Green FM is green-fluorescent mitochondrial stain which appears to localize to mitochondria regardless of mitochondrial membrane potential. The dye will stain live cells but is not well-retained after aldehyde fixation.