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    cox-iv-antibody-mitochondrial-loading-control-ab16056.pdf

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

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  • SDS
Reviews (32)Q&A (19)References (185)

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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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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: WB, ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Xenopus laevis, Chimpanzee, Monkey, Potato, African green monkey, Chinese hamster
  • 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).

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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.

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 (5)
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

  • SDS download

  • Datasheet download

    Download

References (185)

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

ab16056 has been referenced in 185 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
  • Saiki I  et al. Functional Expression of Choline Transporter-Like Protein 1 in LNCaP Prostate Cancer Cells: A Novel Molecular Target. Biomol Ther (Seoul) 28:195-201 (2020). PubMed: 31693854
  • Dibe HA  et al. Epinephrine responsiveness is reduced in livers from trained mice. Physiol Rep 8:e14370 (2020). PubMed: 32061187
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-10 of 32 Abreviews

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-COX IV antibody - Mitochondrial Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Mar 09 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-COX IV antibody - Mitochondrial Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (heart)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization
No
Specification
heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Mar 09 2021

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

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

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