Immunoprecipitation of Human/Mouse Cytochrome c.
Cytochrome c was immunoprecipitated from lysates (5 x 106 cells) of Jurkat human acute T cell leukemia and CTLL‑2 mouse cytotoxic T cell line following incubation with 5 µg Mouse Anti-Human/Mouse/Rat Cytochrome c Monoclonal Antibody (Catalog # MAB899) for 1 hour at 4 °C. Cytochrome c-antibody complexes were absorbed using Protein A Immunoprecipitin (Life Technologies). Immunoprecipitated Cytochrome c (left panel) was detected by Western blot using 0.5 µg/mL Mouse Anti-Human/Mouse/Rat Cytochrome c Monoclonal Antibody (Catalog # MAB897). For additional reference, Western blot (right panel) shows lysates of Jurkat and CTLL2 cell lines probed with 15 µg/mL Mouse Anti-Human/ Mouse/Rat Cytochrome c Monoclonal Antibody (Catalog # MAB897). View our recommended buffer recipes for immunoprecipitation.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Cytochrome c
Cytochrome c is a critical mitochondrial outer
membrane-associated protein in the electron transport chain (1). Reduction and oxidation of an iron molecule
(Fe3+ to Fe2+ and back) within its central heme group
allow it to receive an electron from the cytochrome c1 subunit of cytochrome
reductase and pass it to cytochrome a within the cytochrome oxidase complex (1). Release of cytochrome c from mitochondria
occurs during initiation of apoptotic pathways, either by mechanisms that
rupture or that do not rupture the mitochondial membrane (2, 3). Once in the cytosol, cytochrome c forms a
complex with procaspase 9, recruiting Apaf-1 (apoptotic protease activating
factor-1) and dATP to form apoptosomes, which activate caspase and ultimately
destroy the cell (2, 3).
Cytochrome c is highly conserved, with 90% or greater amino acid
identity among human, mouse, rat, cow and dog sequences (4). After cytochrome c translation, the initial
methionine is cleaved to form the 104 amino acid mature form.
Zubay, G. L., 1983, “Biochemistry”, pp. 380, Addison-Wesley Pub. Co, Inc.
Liu, X. et al., 1996, Cell 86:147.
Li, P. et al., 1997, Cell 91:479.
Matsubara, H and E. L. Smith, 1963, J. Biol. Chem. 238:2732.
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