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
  • Host species
    Mouse
  • 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.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

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

  • Ab33985 staining COX IV in HeLa (Human cervix adenocarcinoma epithelial cell) cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:1000 dilution (1µg/ml). An Alexa Fluor®594 Goat anti-mouse (ab150120) was used as a secondary antibody at 1:1000 dilution (2 µg/ml). Cells were counterstained with anti-Cyclophilin F (ab231155, 5.5µg/ml) and AlexaFluor®488 Goat anti-Rabbit (ab150077, 2µg/ml).  DAPI was used as a nuclear counterstain. Ab33985 was used for negative control 1 at 1:1000 dilution (1µg/ml). For negative control 2, ab231155 was used at a 1:100 dilution (5.5µg/ml) and ab150129 was used as a secondary antibody at 1:1000 dilution (2µg/ml). Confocal image showing mitochondrial staining in HeLa cell line. 

  • 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
    All lanes : 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
    All lanes : 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

    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

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

    All lanes : cow liver membrane with Milk

    Lysates/proteins at 15 µg per lane.

    Blocking peptides at 1.5 % per lane.

    Secondary
    All lanes : Goat Anti-mouse IgG HRP at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa


    Exposure time: 17 hours

    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
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
Username

Abcam user community

Verified customer

Submitted Dec 04 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Loading amount
1e+006 cells
Specification
HEK293
Gel Running Conditions
Reduced Denaturing (4-20)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Mr. Polina Lishko

Verified customer

Submitted Feb 12 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Xenopus laevis Cell lysate - whole cell (egg extract)
Loading amount
15 µg
Specification
egg extract
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Anne-Lore Schlaitz

Verified customer

Submitted Aug 21 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (neuroblastoma cell lines)
Loading amount
10 µg
Specification
neuroblastoma cell lines
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 Feb 11 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
African Green Monkey Cell (COS-1)
Specification
COS-1
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 24 2008

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-1)
Loading amount
20 µg
Specification
COS-1
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 24 2008

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 (kidney)
Specification
kidney
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 14 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human proximal tubular epithelial cell line)
Specification
human proximal tubular epithelial cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X-100 (PBS)
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 14 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (293T and U2OS cells)
Loading amount
30 µg
Specification
293T and U2OS cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Nov 12 2007

21-29 of 29 Abreviews or Q&A

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