Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
- Datasheet
- References (129)
- Protocols
Overview
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Product nameAnti-COX IV antibody - Mitochondrial Loading Control
See all COX IV primary antibodies -
DescriptionRabbit polyclonal to COX IV - Mitochondrial Loading Control
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Host speciesRabbit
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Tested applicationsSuitable for: IHC-Fr, ICC, WB, ICC/IF, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Human, Xenopus laevis, Potato, African green monkey, Chinese hamster
Predicted to work with: Chimpanzee
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human COX IV.
Read Abcam's proprietary immunogen policy (Peptide available as ab16381.) -
Positive control
- This antibody gave a positive signal when tested against: (WB) HeLa and Human Skeletal Muscle Tissue; (IHC-P) Human Liver Tissue; (IF) HeLa.
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General notes
This antibody makes an effective loading control for mitochondria. COX IV is generally expressed at a consistent high level. However, be aware that many proteins run at the same 16kD size as COX IV - our VDAC1 / Porin antibody makes a good alternative mitochondrial loading control for proteins of this size. Some caution is required when using this antibody as a loading control as COXIV expression can vary under some manipulations. An alternative mitochondrial loading control is Mouse monoclonal to COX IV antibody [20E8] (ab14744).
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferpH: 7.40
Preservative: 0.01% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
PurityImmunogen affinity purified
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Primary antibody notesThis antibody makes an effective loading control for mitochondria. COX IV is generally expressed at a consistent high level. However, be aware that many proteins run at the same 16kD size as COX IV - our VDAC1 / Porin antibody makes a good alternative mitochondrial loading control for proteins of this size. Some caution is required when using this antibody as a loading control as COXIV expression can vary under some manipulations. An alternative mitochondrial loading control is Mouse monoclonal to COX IV antibody [20E8] (ab14744).
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab16056 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-Fr | 1/200. | |
| ICC | Use at an assay dependent concentration. | |
| WB | Use a concentration of 0.5 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 17 kDa). | |
| ICC/IF | Use a concentration of 1 µg/ml. | |
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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FunctionThis protein is one of the nuclear-coded polypeptide chains of cytochrome c oxidase, the terminal oxidase in mitochondrial electron transport.
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Tissue specificityUbiquitous.
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Sequence similaritiesBelongs to the cytochrome c oxidase IV family.
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Cellular localizationMitochondrion inner membrane.
- Information by UniProt
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Database links
- Entrez Gene: 454391 Chimpanzee
- Entrez Gene: 1327 Human
- Entrez Gene: 12857 Mouse
- Entrez Gene: 29445 Rat
- Omim: 123864 Human
- SwissProt: O46577 Chimpanzee
- SwissProt: P13073 Human
- SwissProt: P19783 Mouse
see all -
Alternative names
- AL024441 antibody
- COX 4 antibody
- COX IV 1 antibody
see all
Images
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Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Human liver tissue lysate - total protein (ab29889)
Lane 3 : Human heart tissue lysate - total protein (ab29431)
Lane 4 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 17 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)This image is courtesy of an anonymous Abreviewab16056 staining COX IV in Mouse heart tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/600 ) for 12 hours at 4°C. A Cy5® donkey anti-rabbit secondary antibody was used as the secondary antibody.
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Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)ab16056 stained in Hela cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab16056 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature
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Immunohistochemistry (Frozen sections) - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)This image is courtesy of an anonymous Abreviewab16056 staining COX IV in Mouse heart tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 5% serum for 1 hour at 21°C. Samples were incubated with primary antibody (1/200) for 12 hours at 4°C. An Alexa Fluour® 594 donkey anti-rabbit secondary antibody was used as the secondary antibody.
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Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 0.38 µg/ml
Lane 1 : HeLa whole cell lysate
Lane 2 : Human skeletal muscle cell lysate
Lane 3 : HeLa whole cell lysate with Human COX IV peptide (ab16381) at 1 µg/ml
Lane 4 : Human skeletal muscle cell lysate with Human COX IV peptide (ab16381) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/10000 dilution
Predicted band size: 17 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)IHC image of COXIV staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16056, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)This image is courtesy of an anonymous Abreview.ab16056 staining COX IV in breast tumour tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% buffered formalin and blocked with 5% serum for 60 minutes at 21°C; antigen retrieval was by heat mediation in a 10mM sodium citrate buffer pH6. Samples were incubated with primary antibody (1/300 in blocking buffer) for 12 hours at 4°C. An Alexa Fluor® 647-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.
Protocols
References
This product has been referenced in:
- Elena-Real CA et al. Cytochrome c speeds up caspase cascade activation by blocking 14-3-3e-dependent Apaf-1 inhibition. Cell Death Dis 9:365 (2018). Read more (PubMed: 29511177) »
- Li N et al. Renal ischemia/reperfusion-induced mitophagy protects against renal dysfunction via Drp1-dependent-pathway. Exp Cell Res 369:27-33 (2018). Read more (PubMed: 29704468) »
