Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
Key features and details
- Rabbit polyclonal to COX IV - Mitochondrial Loading Control
- Suitable for: WB, ICC/IF, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-COX IV antibody - Mitochondrial Loading Control
See all COX IV primary antibodies -
Description
Rabbit polyclonal to COX IV - Mitochondrial Loading Control -
Host species
Rabbit -
Tested Applications & Species
Application Species ICC/IF HumanIHC-P HumanWB MouseHuman -
Immunogen
Synthetic peptide corresponding to Human COX IV aa 150 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab16381) -
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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Form
Liquid -
Storage instructions
Shipped 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. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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...
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Purity
Immunogen affinity purified -
Primary antibody 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). -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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ICC/IF |
Human
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IHC-P |
Human
|
WB |
Mouse
Human
|
All applications |
Xenopus laevis
Chimpanzee
Monkey
Potato
African green monkey
Chinese hamster
|
Application | Abreviews | Notes |
---|---|---|
WB | (15) |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 17 kDa).
|
ICC/IF | (5) |
Use a concentration of 1 µg/ml.
|
IHC-P | (3) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
Notes |
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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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Function
This protein is one of the nuclear-coded polypeptide chains of cytochrome c oxidase, the terminal oxidase in mitochondrial electron transport. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the cytochrome c oxidase IV family. -
Cellular localization
Mitochondrion 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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All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml
Lane 1 : Human skeletal muscle tissue lysate
Lane 2 : Rat skeletal muscle tissue
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 17 kDa
Observed band size: 17 kDaBlocking buffer: 2% BSA
Gel type: MES
Exposure Time: 1 minute
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Lane 1 : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml (Human skeletal muscle tissue lysate)
Lanes 2-3 : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml
Lane 2 : Human heart tissue lysate
Lane 3 : Mouse skeletal muscle tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 17 kDa
Observed band size: 17 kDaBlocking buffer: 2% BSA
Gel type: MES
Exposure Time: 1 minute
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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 Abreview
ab16056 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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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.
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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 (174)
ab16056 has been referenced in 174 publications.
- Ghazi S et al. Multiparametric imaging reveals that mitochondria-rich intercalated cells in the kidney collecting duct have a very high glycolytic capacity. FASEB J 34:8510-8525 (2020). PubMed: 32367531
- Schlaak RA et al. Differences in Expression of Mitochondrial Complexes Due to Genetic Variants May Alter Sensitivity to Radiation-Induced Cardiac Dysfunction. Front Cardiovasc Med 7:23 (2020). PubMed: 32195269
- Zhang XH et al. Heat shock protein 90 relieves heat stress damage of myocardial cells by regulating Akt and PKM2 signaling in vivo. Int J Mol Med 45:1888-1908 (2020). PubMed: 32236591
- Chuang YC et al. Sirtuin 1 Regulates Mitochondrial Biogenesis and Provides an Endogenous Neuroprotective Mechanism Against Seizure-Induced Neuronal Cell Death in the Hippocampus Following Status Epilepticus. Int J Mol Sci 20:N/A (2019). PubMed: 31340436
- Arrázola MS et al. Axonal Degeneration Is Mediated by Necroptosis Activation. J Neurosci 39:3832-3844 (2019). PubMed: 30850513