DePsipherTM Mitochondrial Potential Assay
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The mitochondrial permeability transition is an important event
in the apoptotic process wherein the electrochemical gradient (referred
to as Dym) across the mitochondrial membrane collapses. In some
apoptotic pathways, the collapse is thought to occur through the
formation of pores in the mitochondria, possibly involving Bax insertion
and dimerization, and is accompanied by the release of cytochrome
c into the cytoplasm.
The DePsipher™ Kit uses a unique cationic dye (5,5’6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzimidazolylcarbocyanine
iodide) to indicate the loss of the mitochondrial potential. The
dye readily enters cells and fluoresces brightly red in its multimeric
form within healthy mitochondria. In apoptotic cells, the mitochondrial
membrane potential collapses, and the DePsipher™ reagent cannot accumulate
within the mitochondria. In these cells, DePsipher™ remains in the
cytoplasm as a green fluorescent monomeric form. Apoptotic cells,
showing primarily green fluorescence, are easily differentiated
from healthy cells which show red fluorescence. The aggregate red
form has absorption/emission maxima of 585/590 nm, and the green
monomeric form has absorption/emission maxima of 510/527 nm. Both
apoptotic and healthy cells can be visualized simultaneously by
epifluorescence microscopy using a wide band-pass filter.
The DePsipher™ reagent is easy to use. Simply resuspend the reagent
in Reaction Buffer or culture media (with or without the Stabilizer
Solution), add to your cells, incubate for 15 to 20 minutes, wash
and analyze by flow cytometry or microscopy. Visualization by microscopy
allows a rapid inspection and qualification of apoptosis. Flow cytometric
analysis allows easy quantitation of cell death as evidenced by
mitochondrial potential breakdown.
| Features: |
Simple. Just add DePsipher™ reagent to media or reaction buffer.
Unique Stabilizer Solution improves results.
Fast. Takes only 20 minutes.
Flexible. View cells by epifluorescence or confocal microscopy, or analyze cells by flow cytometry.
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| Applications: |
Flow cytometry
Epifluorescence microscopy
Confocal microscopy
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| Components: |
Part # |
Component |
Size |
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6300-100-01 |
DePsipher |
100 µl |
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6300-100-02 |
10X Reaction Buffer |
2 x 30ml |
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6300-100-03 |
Stabilizer Solution |
5ml |
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| Storage: |
Components are stored at -20°C and 4°C.
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