Cytochrome c is a requisite component of the mitochondrial respiratory chain and participates in the electron transfer between complex III and complex IV of the respiratory chain. Cytochrome c is a soluble protein that is localized in the mitochondrial intermembrane space and is loosely attached to the surface of the inner membrane. The protein is synthesized on cytoplasmic ribosomes as apocytochrome c and then translocated into the mitochondria via a unique pathway independent of the general protein translocation machinery, protease-sensitive components of the outer membrane, or a membrane potential across the inner membrane. In response to a variety of apoptotic stimuli, Cytochrome c is released from mitochondria into the cytosol. This release can be blocked by Bcl-2 and suggests that Bcl-2 may function by regulating Cytochrome c release. Cytosolic Cytochrome c is an essential component of the vertebrate apoptosome which is composed of Cytochrome c, Apaf-1, and procaspase-9 (Apaf-3). Interaction of these proteins leads to the activation of caspase-9, which in turn activates other caspases (such as caspase-3) ultimately resulting in cell death. The current working hypothesis is that release of Cytochrome c from mitochondria commits a cell to die either by a “rapid” apoptotic mechanism involving activation of the Apaf-1/caspase-9 cascade, or by a “slower” necrotic process involving the collapse of the electron transport chain. ||Applications: |Suitable for use in Immunofluorescence, ELISA and Immunoprecipitation. Other applications not tested.||Recommended Dilution:|ELISA (Native): 0.1-1ug/ml |Immunofluorescence: 20ug/ml |Immunoprecipitation (Native): 15ug (for extract from ~10e6 cells) |Optimal dilutions to be determined by the researcher.|Positive Control:|HeLa, Rat6||Storage and Stability:|Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Note: Sodium azide is a potent inhibitor of peroxidase and should not be added to HRP conjugates. |For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.||Note: Applications are based on unconjugated antibody.