Anti-COX IV antibody (ab66739)
- Product nameAnti-COX IV antibodySee all COX IV primary antibodies ...
- DescriptionRabbit polyclonal to COX IV
- Tested applicationsWB, IHC-P, ICC/IF more details
- Species reactivityReacts with: Human
Predicted to work with: Rat, Chicken, Cow, Dog, Pig, Chimpanzee
A region within synthetic peptide: AISTSVCVRA HESVVKSEDF SLPAYMDRRD HPLPEVAHVK HLSASQKALK, corresponding to amino acids 14-63 of Human COX IV
- Positive controlHepG2 cell lysate. Human heart tissue. Human kidney tissue. This antibody gave a positive result when used in the following methanol fixed cell lines: mcf-7.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPreservative: None
Constituents: 2% Sucrose, PBS
- Concentration information loading...
- PurityProtein A purified
- Clonality Polyclonal
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Integration of energy
Our Abpromise guarantee covers the use of ab66739 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use at a concentration of 1.25 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 20 kDa).
ELISA titre using peptide based assay: 1/312500.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
- FunctionThis protein is one of the nuclear-coded polypeptide chains of cytochrome c oxidase, the terminal oxidase in mitochondrial electron transport.
- Tissue specificityUbiquitous.
- Sequence similaritiesBelongs to the cytochrome c oxidase IV family.
- Cellular localizationMitochondrion inner membrane.
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Anti-COX IV antibody images
ICC/IF image of ab66739 stained MCF-7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab66739 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Anti-COX IV antibody (ab66739) at 1.25 µg/ml + HepG2 cell lysate at 10 µg
HRP conjugated anti-Rabbit IgG at 1/50000 dilution
Predicted band size : 20 kDa
Observed band size : <21 kDa (why is the actual band size different from the predicted?)
Antibody diltued in 5% skim milk / PBS buffer. Gel concentration 12%.
ab66739, at 2 µg/ml, staining COX IV in Human heart tissue by immunohistochemistry on paraffin-embedded tissue. The arrows indicate positively stained myocardial cells.
ab66739, at 2 µg/ml, staining COX IV in human kidney tissue by immunohistochemistry on paraffin-embedded tissue. Positively stained cells are epithelial cells of the renal tubule.
All lanes : Anti-COX IV antibody (ab66739)
Lane 1 : Lysate prepared from human liver
Lane 2 : Lysate prepared from rat liver
Lane 3 : Lysate prepared from wild type mouse liver
Lane 4 : Lysate prepared from AMPKa1+2-/- mouse liver
Lane 5 : Lysate prepared from human muscle
Lane 6 : Lysate prepared from rat muscle
Lane 7 : Lysate prepared from mouse muscle
Predicted band size : 20 kDa
A small piece (~30mg) of every organ was homogenized in a 6:1 (v/w) ratio of ice-cold buffer containing: 50 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (pH 7.6), 50 mM sodium fluoride, 50 mM potassium chloride, 5 mM sodium pyrophosphate, 1 mM ethylenediaminetetraacetic acid, 1 mM ethylene glycol tetraacetic acid, 5 mM ß-glycerophosphate, 1 mM sodium vanadate, 1 mM dithiothreitol, 1% nonyl phenoxypolyethoxylethanol (Tergitol-type NP40) and protease inhibitors cocktail. Homogenates were centrifuged (13,200 rpm; 15 min, 4°C) and the protein content of the supernatant was determined using a bicinchoninic acid (BCA) protein assay kit.Proteins (30 µg) were separated by either 12.5 or 10% SDS-PAGE followed by transfer to a polyvinylidene fluoride transfer membrane. Membranes were blocked for 1 h at room temperature in tris-buffered saline tween-20 buffer with 5% non-fat dry milk followed by an overnight incubation with antibodies. Blots were then incubated with horseradish