Monoclonal Antibody to Cytochrome C (Clone 7H8.2C12) Biotin Conjugate

Background
In addition to its well-known role in electron transport, cytochrome c (CYT) has been identified as a mediator of apoptosis or programmed cell death (1). Cytochrome c translocates from mitochondria into the cytoplasm in the early phases of apoptosis and plays a role in initiation of the apoptotic proteolytic cascade by activating caspase-3 (2). Cytochrome c has been shown to bind Apaf-1. Apaf-1 has also been shown to activate capase-3 (3).

Antigen
Synthetic peptides corresponding to amino acids 1-80, 81-104 and 66-104 of pigeon CYT were used as the immunogen (Jemmerson et al. 1991). The antibody recognizes an epitope within amino acids 93-104 of pigeon Cytochrome c. based on competitive ELISA results (Jemmerson et al. 1991).

Application Notes
1. An ~15 kDa band is observed. 2. This antibody recognizes total cytochrome C which includes both apocytochrome c (i.e. cytochrome c in the cytosol without heme attached) and holocytochrome c (i.e. cytochrome c in the mitochondria with heme attached).

Reference
1. Fernandes-Alnemri, T., Litwack, G., and Alnemri, ES. CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1 beta-converting enzyme J. Biol. Chem. 269: 30761-30764 (1994). PMID: 7983002.
2. Liu, X., Kim, CN, Yang, J., Jemmersen, R., and Wang, X. Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 86: 147-157 (1996). PMID: 8689682.
3. Jemmersen, R. Johnson, J.G., Burell, E., Taylor, P.S., and Jemkins, M.K. A monoclonal antibody specific for a cytochrome c T cell stimulatory peptide inhibits T cell response and affects the way the peptide associates with antigen-presenting cells. Eur. J. Immunol. 21: 143-151 (1991). PMID: 1846813.
4. Zou, H., Henzel, WJ, Liu, X., Lutschg, A., and Wang, X. Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell 90: 405-413 (1997). PMID: 9267021.