Mitochondrial & Cytochrome c Apoptosis Kits

Disruption of mitochondrial transmembrane potential is one of the earliest intracellular events that occurs upon induction of apoptosis. The Mitochondrial Apoptosis Staining Kit provides a simple and sensitive, fluorescent-based in vitro assay for detecting the mitochondrial changes in apoptosis and distinguishing between healthy and apoptotic cells. The assay utilizes a cationic dye (called MitoStain) that fluoresces differently in healthy versus apoptotic cells. In healthy cells, MitoStain accumulates and aggregates in the mitochondria - fluorescing brightly red. In apoptotic cells, a shift in mitochondrial transmembrane potential prevents the dye from accumulating in the mitochondria. Instead it remains in the cytoplasm in its monomer form - fluorescing green. The fluorescent signals can be easily detected by fluorescence microscopy or flow cytometry using common filters and channels, respectively (FITC, rhodamine, PI). The assay takes approximately 20 minutes, is highly sensitive and detects apoptosis reliably in living cells.

Cytochrome c has been shown to play an important role in apoptosis. The protein is located in the space between the inner and outer mitochondrial membranes. An apoptotic stimulus triggers the release of cytochrome c from the mitochondria into the cytosol where it binds to Apaf-1. The cytochrome c/Apaf-1 complex activates caspase-9 which then activates caspase-3 and other downstream caspases of the apoptotic cascade. The PromoKine Cytochrome c Apoptosis Detection Kit provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis. The kit provides an easy-to-use procedure and unique reagents for separating a highly enriched mitochondria fraction from cytosol. Cytochrome c released from mitochondria into cytosol is then determined by Western blotting using the cytochrome c antibody provided in the kit. The procedure is very simple and straightforward - no ultracentrifugation is required and no toxic chemicals are involved.