MitoLight Mitochondrial Apoptosis Detection Kit

Disruption of the mitochondrial transmembrane potential is one of the earliest intracellular events that occur following induction of apoptosis. The MitoLight Apoptosis Detection Kit utilizes a lipophilic cation, termed as MitoLight, as a mitochondrial activity marker. MitoLight is a mitochondrial dye that stains mitochondria in living cells in a membrane potential-dependent fashion. The monomer is in equilibrium with so-called J-aggregates, which are favored at higher dye concentration or higher mitochondrial membrane potential.

MitoLight partitions differently in healthy cells than in apoptotic cells. Therefore, it has been possible to use a fluorescence ratioing technique to study mitochondrial membrane potentials. In healthy cells, the dye accumulates and aggregates in the mitochondria, giving off a bright red fluorescence (λem = 585-590 nm). In apoptotic cells with altered mitochondrial membrane potential, the dye in its monomeric form stays in the cytoplasm, fluorescing green (λem = 527-530 nm), providing a ready discrimination between apoptotic and nonapoptotic cells. The fluorescence can be observed by fluorescence microscopy using a band-pass filter (detects FITC and rhodamine) or analyzed by flow cytometry using FITC channel for green monomers (Ex/Em = 488/530) and PI channel for red aggregates (Ex/Em = 488/585).

Research Category
Apoptosis & Cancer

The MitoLight Apoptosis Detection Kit for flow cytometry utilizes a lipophilic cation, termed as MitoLight, as a mitochondrial activity marker.

Store kit materials at -20°C up to their expiration date. After thawing, unused kit components can be refrozen. For best results, aliquot the MitoLight reagent into separate vials to avoid repeated freeze/thaw cycles.

PROTECT REAGENTS FROM LIGHT.

MitoLight (Part No. 71616): 100 μL in DMSO.

10X Incubation Buffer (Part No. 71614): 20 mL.

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