Cytochrome C, Rat, Western Blot Positive Control

Cytochrome-C enzyme is a nuclear encoded protein located in the mitochondria of all aerobic cells. It is involved in the electron transport system that functions in oxidative phosphorylation. It accepts electrons from cytochrome b and transfers them to cytochrome oxidase. In the process the iron of the heme group (which is identical to that of hemoglobin and myoglobin) shifts from the ferrous to the ferric state. Human cytochrome-c has 105-aa (~12kD). Mitochondria play a pivotal role in the regulation of programmed cell death or apoptosis. Apoptosis is driven by two classes of specialized proteases known as caspases (Cysteine aspartase). The initiator caspases can be activated by self-cleavage. The effector caspases are then activated in an amplification cascade. Several key participants are released from the mitochondria that regulate apoptosis. The first such factor (Cytochrome-C) to be described binds to a cytoplasmic scaffolding protein called Apaf-1. Binding of the mitochondria factor allows Apaf-1 to form a ternary complex with, and activate, the initiator pro-caspase-9. Active caspase-9 then turns on downstream effector caspases, initiating apoptosis. Murine embryos devoid of cytochrome c died in utero by midgestation. In contrast, cells lacking cytochrome c demonstrated increased sensitivity to cell death signals triggered by TNF. , Aapplications:, Suitable for use in Western Blot. Other applications not tested. Recommended Dilution: Western Blotting Load ~10 ul/lane to visualize with appropriate antibodies. , Optimal dilution determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.