Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)

Overview

  • Product name
    Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker
    See all COX IV primary antibodies
  • Description
    Mouse monoclonal [mAbcam33985] to COX IV - Mitochondrial Marker
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-Fr, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Cow, Human, Xenopus laevis, Monkey, African green monkey, Chinese hamster, Drosophila C virus
    Predicted to work with: Chimpanzee, Zebrafish
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human COX IV aa 150 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab16381)

  • Positive control
    • WB: Jurkat and HepG2 whole cell lysates and human skeletal muscle, mouse skeletal muscle and cow kidney tissue lysates.
  • General notes

    This antibody clone is manufactured by Abcam.

Properties

Applications

Our Abpromise guarantee covers the use of ab33985 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
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use at an assay dependent concentration.

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 [mAbcam33985] - Mitochondrial Marker (ab33985) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 4 : Skeletal Muscle (Mouse) Tissue Lysate
    Lane 5 : Kidney (Cow) Tissue Lysate (ab29073)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 15 kDa


    Exposure time : 1 minute
  • ICC/IF image of ab33985 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33985, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • Overlay histogram showing HeLa cells stained with ab33985 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33985, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution

    Lane 1 : Crude extract prepared from Xenopus laevis egg
    Lane 2 : Cytosol lysate prepared from Xenopus laevis egg extract
    Lane 3 : Total membrane lysate prepared from Xenopus laevis egg extract

    Lysates/proteins at 15 µg per lane.

    Secondary
    HRP conjugated donkey anti-mouse IgG at 1/4000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 15 kDa


    Exposure time : 90 minutes

    This image is courtesy of an Abreview submitted by Dr Anne-Lore Schlaitz

    See Abreview

  • ab33985 staining COX IV in human proximal tubular epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 in PBS and blocked with 3% BSA for 15 minutes at 20°C. Samples were incubated with primary antibody (1/200 in PBS) for 45 minutes at 20°C. ab6785, a FITC-conjugated goat anti-mouse IgG (H+L) polyclonal was used as the secondary antibody (1/1000).

    See Abreview

  • ab33985 staining COX IV in mouse kidney (tubules) tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized with 0.2% trition X-100 and blocked with 5% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in sodium citrate buffer pH 6. Samples were incubated with primary antibody (1/200 in PBS) for 9 hours at 4°C. An Alexa Fluor® 594-conjugated goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. DAPI was used for staining the nucleus.

    See Abreview

References

This product has been referenced in:

See all 28 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (leukocytes from bone marrow)
Specification
leukocytes from bone marrow
Fixative
Methanol
Permeabilization
No
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Jan 27 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Tubules (Kidney))
Specification
Tubules (Kidney)
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer pH 6.0
Permeabilization
Yes - 0.2% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Dec 30 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Macrophages, Fibroblasts, PBMC)
Loading amount
20 µg
Specification
Macrophages, Fibroblasts, PBMC
Gel Running Conditions
Reduced Denaturing
Blocking step
Special fluorescence blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: RT°C
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Verified customer

Submitted Oct 13 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Brain lysate)
Loading amount
50 µg
Specification
Brain lysate
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Aug 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293 FT cell line)
Loading amount
50 µg
Specification
293 FT cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Aug 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
African Green Monkey Cell (COS cell line)
Specification
COS cell line
Fixative
Acetone
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Aug 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (COS Cell line)
Loading amount
50 µg
Specification
COS Cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (bone marrow cells)
Specification
bone marrow cells
Fixative
Formaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted May 10 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (293FT)
Specification
293FT
Fixative
Formaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Jan 14 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Liver)
Specification
Liver
Fixative
Formaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
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Verified customer

Submitted Dec 04 2009

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