Overview

  • 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
    Suitable for: IHC-Fr, ICC, WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis, Potato, African green monkey, Chinese hamster
    Predicted to work with: Chimpanzee
  • 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.
  • 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

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

  • 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
  • Database links
  • Alternative names
    • AL024441 antibody
    • COX 4 antibody
    • COX IV 1 antibody
    • COX IV antibody
    • COX IV-1 antibody
    • Cox4 antibody
    • COX41_HUMAN antibody
    • Cox4a antibody
    • COX4B antibody
    • COX4I1 antibody
    • COX4I2 antibody
    • COX4L2 antibody
    • COXIV antibody
    • Cytochrome c oxidase polypeptide IV antibody
    • Cytochrome c oxidase subunit 4 isoform 1 mitochondrial antibody
    • Cytochrome c oxidase subunit 4 isoform 1, mitochondrial antibody
    • Cytochrome C Oxidase subunit IV antibody
    • Cytochrome c oxidase subunit IV isoform 1 antibody
    • Cytochrome c oxidase subunit IV isoform 2 (lung) antibody
    • Cytochrome c oxydase subunit 4 antibody
    • dJ857M17.2 antibody
    • MGC105470 antibody
    • MGC72016 antibody
    see all

Images

  • 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

  • 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.

    See Abreview

  • 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

  • ab16056 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.

    See Abreview

  • 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?

  • 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.
  • 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.

    See Abreview

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) »
See all 129 Publications for this product

Customer reviews and Q&As

1-10 of 47 Abreviews or Q&A

Answer

Thank you very much for your feedback.

We would like to encourage you to submit an Abreview about your experiments with ab16056 in drosophila. This information is very valuable for our customers when choosing an antibody.

You would earn Abpoints that can be redeemed asdiscount on a future purchase or Amazon gift vouchers.

We are looking forward to your Abreview and wish you continuing luck with your research

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (heart)
Permeabilization
Yes - 0.2% triton X
Specification
heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 07 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (heart)
Gel Running Conditions
Non-reduced Denaturing (15%)
Loading amount
20 µg
Specification
heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Sep 07 2018

Answer

The antibodies detect different subunits of cytochrome c oxidase, which is also known as Complex IV, and which is abbreviated as CIV on the ab110413 datasheet (Total OXPHOS Rodent WB Antibody Cocktail).

The CIV (Complex IV) antibody in the panel ab110413 detects Compex IV subunit 1, also called COXI and MT-CO1, at ˜40kDa. This antibody is available separately as ab14705. The COX IV mitochondria loading control antibody ab16056 detects Complex IV subunit 4, also known as COXIV, at about 15kDa. Here are links to the UniProt records for the human proteins.

MT-CO1 https://www.uniprot.org/uniprot/P00395
COXIV https://www.uniprot.org/uniprot/P13073

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (SH-SY5Y)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
SH-SY5Y
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Mr. Chun Chau SUNG

Verified customer

Submitted May 23 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
African green monkey Cell (BS-C-1 cells)
Permeabilization
Yes - 0.2% Triton X
Specification
BS-C-1 cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Mr. J. Y. Seo

Verified customer

Submitted Apr 11 2018

Application
Western blot
Sample
Pig Cell lysate - other (Porcine kidney epithelial cells (PK-15))
Gel Running Conditions
Reduced Denaturing (4-20% Tris-Glycine)
Loading amount
10 µg
Treatment
100 μg/ml poly(I:C) for 18 hrs
Specification
Porcine kidney epithelial cells (PK-15)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Mr. Sam Hardy

Verified customer

Submitted Jan 16 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (carotid body)
Permeabilization
Yes - 0.5% TritonX in PBS
Specification
carotid body
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
4% PFA

Abcam user community

Verified customer

Submitted Oct 06 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Bone marrow)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization
No
Specification
Bone marrow
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Mar 08 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (breast tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium citrate buffer pH6
Permeabilization
No
Specification
breast tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% buffered formalin

Abcam user community

Verified customer

Submitted Mar 07 2016

1-10 of 47 Abreviews or Q&A

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