For best results with this antibody in Western blot, do not boil samples before loading onto the gel. Boiling of the sample will cause a loss of signal. Hydrophobic intrinsic membrane proteins such as the core mtDNA-encoded proteins of the mitochondrial OXPHOS complexes tend to run faster in SDS-PAGE than predicted by their amino acid composition. This is likely due to incomplete unfolding of the protein and a more negative charge:mass ratio. In Western blot applications, this antibody detects a band at approximately 40 kDa (predicted MW: 57 kDa). In mouse liver lysate a specific band below 37 kDa was detected.
Recommended positive controls:
ICC/IF - Pig retinal pigment epithelial cells, rat cerebellum primary cells
IHC (P) - Human skeletal muscle and colon tissue, rat pancreas tissue, pig smooth muscle tissue
WB - Mouse, human, bovine and rat heart mitochondria lysate
Flow - HEK-293 cells
Cytochrome c oxidase subunit I (CO1 or MTCO1) is 1 of 3 mitochondrial DNA (mtDNA) encoded subunits (MTCO1, MTCO2, MTCO3) of respiratory Complex IV. Complex IV is located within the mitochondrial inner membrane and is the third and final enzyme of the electron transport chain of mitochondrial oxidative phosphorylation. It is composed of 13 polypeptides. Subunits I, II, and III (MTCO1, MTCO2, MTCO3) are encoded by mtDNA while subunits IV, Va, Vb, VIa, VIb, VIc, VIIa, VIIb, VIIc, and VIII are nuclear encoded. The cytochrome c oxidase family of enzymes have 4 redox centers, 2 hemes and 2 copper centers. In mitochondrial Complex IV, the 2 hemes are a and a3 and the 2 coppers are CuA and CuB. The 2 hemes and CuB are bound to subunit I. Acin-Perez et al. (2003) identified a cell line containing single and double missense mutations in the cytochrome c oxidase (COX) subunit I gene of mouse mitochondrial DNA. And they hypothesized that deleterious mutations can arise and become predominant; cultured cells can maintain several mtDNA haplotypes at stable frequencies; the respiratory chain has little spare COX capacity; and that the size of a cavity in the vicinity of val421 in MTCO1I of animal COX may affect the function of the enzyme.