Cytochrome c (cyt-c) plays 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 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. Cytochrome c Releasing Apoptosis Assay Kit provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis. The kit provides unique formulations of reagents to isolate a highly enriched mitochondria fraction from cytosol. The procedure is so simple and easy to perform, no ultracentrifugation is required and no toxic chemicals are involved. Cytochrome c releasing from mitochondria into cytosol is then determined by Western blotting using the cytochrome c antibody provided in the kit. Kit Contents (100 assays): VolumeCapStorageA2299-90A: Mitochondria Extraction Buffer 10mlNM 4°C A2299-90B: 5X Cytosol Extraction Buffer 20mlWM 4°CA2299-90C: DTT (1M) 110ulBlue -20°CA2299-90D: 500X Protease Inhibitor Cocktail(Add 250ml of DMSO before use. 1 vialRed -20°CA2299-90E: Cytochrome c Antibody 500ul Green -20°C
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