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): Volume Cap Storage
A2299-90A: Mitochondria Extraction Buffer 10ml NM 4 degrees C
A2299-90B: 5X Cytosol Extraction Buffer 20ml WM 4 degrees C
A2299-90C: DTT (1M) 110ul Blue -20 degrees C
A2299-90D: 500X Protease Inhibitor Cocktail
(Add 250ml of DMSO before use. 1 vial Red -20 degrees C
A2299-90E: Cytochrome c Antibody
(0.2mg/ml) 500ul Green -20 degrees C