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Tags & Cell Markers Subcellular Markers Organelles Mitochondria
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Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)

  • Datasheet
  • SDS
Reviews (30)Q&A (19)References (174)

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Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
  • Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
  • Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
  • Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

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Protein
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Recombinant Human COX IV protein (ab112273)
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Alexa Fluor® 488 Anti-MTCO1 antibody [1D6E1A8] (ab154477)

View more associated products

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 & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human COX IV aa 150 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab16381)

  • 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

  • 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...
  • 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

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Mitochondria
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Isotype/Loading Controls
    • Loading Controls
    • COX IV - Mitochondrial
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Oxidative phosphorylation
    • Complex IV
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human COX IV protein (ab112273)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
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
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.
  • Target information above from: UniProt accession P13073 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • SwissProt: P10888 Rat
    • Unigene: 433419 Human
    • Unigene: 386758 Mouse
    • Unigene: 2528 Rat
    see all
  • 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

  • Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
    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 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 kDa



    Blocking buffer: 2% BSA

    Gel type: MES

    Exposure Time: 1 minute

  • Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
    Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
    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 kDa



    Blocking buffer: 2% BSA

    Gel type: MES

    Exposure Time: 1 minute

  • Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
    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)
    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.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
    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

  • Western blot - Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)
    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)
    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)
    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.

    See Abreview

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (174)

    Publishing research using ab16056? Please let us know so that we can cite the reference in this datasheet.

    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
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 49 Abreviews or Q&A

    Question

    Inquiry: I have the answer on one of the question for the Ab16056 namely "Are these antibodies tested with Drosophila species?" . I've used this antibody for immunofluorescence staining in S2 cells yesterday and I have got pictures showing this Ab works very well in Drosophila. Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on May 08 2012

    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

    Abcam Scientific Support

    Answered on May 08 2012

    Immunocytochemistry abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Rat Cultured Cells (Cardiomyocytes)
    Permeabilization
    Yes - 0.02% Triton-X-100
    Specification
    Cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted May 07 2019

    Immunocytochemistry abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (HEK293)
    Permeabilization
    Yes - 0.02% Triton-X-100
    Specification
    HEK293
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted May 07 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 07 2018

    Western blot abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 07 2018

    Question

    Hi, this antibody panel, ab110413, recognizes complex I-V quite sufficiently. I was wondering if there is any difference between the MTCO1 (COX IV subunit I) at 40kDa and another COX IV loading control (ab16056) at 17kDa? Since COX IV is typically used for loading control in the mitochondrial western blot. I use that Ab for other mitochondrial protein analysis. But I am not sure if I can use COX IV in the case of evaluating MTCO1, since they are in some way the same COX IV, right? And why the molecular weight for the proteins recognized by both Abs are different? Thanks! Hailong

    Read More

    Abcam community

    Verified customer

    Asked on Jul 16 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

    Abcam Scientific Support

    Answered on Jul 16 2018

    Western blot abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mr. Chun Chau SUNG

    Verified customer

    Submitted May 23 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mr. J. Y. Seo

    Verified customer

    Submitted Apr 11 2018

    Western blot abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mr. Sam Hardy

    Verified customer

    Submitted Jan 16 2017

    Immunohistochemistry (Frozen sections) abreview for Anti-COX IV antibody - Mitochondrial Loading Control

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 06 2016

    1-10 of 49 Abreviews or Q&A

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