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Mitochondrial Membrane Potential Diruption Assay
Disruption of the electrochemical gradient across the mitochondrial transmembrane is one of the early intracellular events to occur following induction of apoptosis. The DePsipher™ kit uses a lipophilic cation to identify changes in the potential of the mitochondrial membrane.
In normal, healthy cells, the DePsipher reagent aggregates in the mitochondria to form an orange fluorescent compound. However, in apoptotic cells, when the mitochondrial membrane potential is disrupted, the DePsipher reagent remains in its monomeric form and fluoresces green. These changes in fluorescence can be monitored either by microscopy or flow cytometry.
Identification of Apoptotic INT407 Cells using the DePsipher Kit. NT407 human embryonic intestinal cells were treated with 25 µM etoposide for 8 hours and then incubated with the DePsipher reagent in reaction bugger for 30 minutes (both provided in the DePsipher Kit, Catalog # 6300-100-K). Healthy cells (containing red aggregates) were differentiated from apoptotic cells (containing only green monomers) by fluorescence microscopy.