Anti-COX IV antibody [20E8C12] (ab14744)

Reviews (27)Q&A (24)References (203)

Overview

  • Product name

    Anti-COX IV antibody [20E8C12]
    See all COX IV primary antibodies

  • Description

    Mouse monoclonal [20E8C12] to COX IV

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, IHC-FoFr, WB, ICC/IF, IHC-Fr, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Hamster, Cow, Human, Pig, Drosophila melanogaster, Zebrafish

  • Immunogen

    Tissue, cells or virus corresponding to Cow COX IV.

  • Positive control

    • Human, bovine, murine and rat heart mitochondria.

  • General notes

    This antibody clone is manufactured by Abcam.

    This antibody makes an effective loading control for mitochondria. COXIV is generally expressed at a consistent high level. However, be aware that many proteins run at the same 16kD size as COXIV - 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 Rabbit polyclonal to COX IV antibody (ab16056).

    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 ab14744 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 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-FoFr Use at an assay dependent concentration. PubMed: 18698340
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IP 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

    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 [20E8C12] (ab14744)
    Western blot - Anti-COX IV antibody [20E8C12] (ab14744)This image is courtesy of an anonymous Abreview
    Anti-COX IV antibody [20E8C12] (ab14744) at 1/1000 dilution + Fruit fly (Drosophila melanogaster) heart tube tissue lysate at 10 µg

    Secondary
    HRP-conjugate Goat anti-mouse IgG polyclonal at 1/4000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 1 minute

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [20E8C12] (ab14744)
    Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [20E8C12] (ab14744)This image is courtesy of an anonymous Abreview

    ab14744 at 1/100 staining MCF-10A cells (human mammary epithelial cell line) by ICC/IF. The cells were methanol/acetone fixed at -20C for 5 minutes, blocked with BSA and then incubated with the antibody for 16 hours. The positive tissue was colocalised with a mitotracker (mitotrackers are a series of patented mt-selective stains that are concentrated by active mt and well retained during cell fixation). An Alexa-Fluor ® 488 conjugated goat anti-mouse antibody was used as the secondary. The image shows COXIV staining in green and DAPI staining in blue.

    See Abreview

  • Flow Cytometry - Anti-COX IV antibody [20E8C12] (ab14744)
    Flow Cytometry - Anti-COX IV antibody [20E8C12] (ab14744)
    Overlay histogram showing HeLa cells stained with ab14744 (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 (ab14744, 0.5µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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.

  • Western blot - Anti-COX IV antibody [20E8C12] (ab14744)
    Western blot - Anti-COX IV antibody [20E8C12] (ab14744)
    All lanes : Anti-COX IV antibody [20E8C12] (ab14744) at 1/5000 dilution

    Lane 1 : Cytoplasmic fraction mouse NIH/3T3 cell lysate
    Lane 2 : Nuclear fraction mouse NIH/3T3 cell lysate

    Secondary
    All lanes : HRP conjugated goat anti-mouse antibody

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 16 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    This image is courtesy of an Abreview submitted by Camilla Skjerpen on 4 July 2005.

    See Abreview

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-COX IV antibody [20E8C12] (ab14744)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-COX IV antibody [20E8C12] (ab14744)Image from Hashimoto T. et. al., PLoS ONE. 2008; 3(8): e2915 (Fig. 1B)

    ab14744 staining COX IV in rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Rats were anesthetized and intracardially perfused with 500 ml of normal saline at room temperature, followed by 500 ml of ice-cold, freshly made 4% paraformaldehyde in phosphate buffer (PB, 0.1 M, pH 7.4). A Cy3 conjugated anti mouse antibody was used as secondary. 

  • Western blot - Anti-COX IV antibody [20E8C12] (ab14744)
    Western blot - Anti-COX IV antibody [20E8C12] (ab14744)
    All lanes : Anti-COX IV antibody [20E8C12] (ab14744) at 1 µg/ml

    Lane 1 : Isolated mitochondria from human heart at 5 µg
    Lane 2 : Isolated mitochondria from bovine heart at 1 µg
    Lane 3 : Isolated mitochondria from rat heart at 10 µg
    Lane 4 : Isolated mitochondria from murine heart at 10 µg

    Observed band size: 16 kDa why is the actual band size different from the predicted?

References

This product has been referenced in:

See all 203 Publications for this product

Publishing research using ab14744? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 51 Abreviews or Q&A

Question

Klone heißen anders/kürzer:
ab14744 : 20E8
ab14705 : 1D6E
Kundin hat diese AKs am 10.1.2011 bestellt, kamen als Lyophilisat und wurden eingefroren bei -20C
Ist das okay?
Sind die Klone dieselben?

Read More
Answer

Vielen Dank für Ihren Anruf.

Ich freue mich Ihnen bestätigen zu können, dass die Klone dieselben sind und früher bei MitoScience nur abgekürzt wurden.

Ich kann Ihnen auch bestätigen, dass eine Lagerung bei -20C korrekt ist solange die Antikörper lyophilisiert sind.

Bitte lagern sie die Antikörper bei +4C nach Rekonstitution.

Read More

Western blot abreview for Anti-COX IV antibody [20E8C12]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Sheep Cell lysate - other (preadipocyte mitochondrial isolates v. cytoplasmic)
Gel Running Conditions
Reduced Denaturing (10% PAG)
Loading amount
10 µg
Specification
preadipocyte mitochondrial isolates v. cytoplasmic
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Miss. Barbara Makela

Verified customer

Submitted Oct 13 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-COX IV antibody [20E8C12]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Chinese Hamster Cell (CHO cell)
Specification
CHO cell
Permeabilization
Yes - 1% TRITON-X-100
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Mar 18 2015

Western blot abreview for Anti-COX IV [20E8C12] antibody

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (HEPATOCYTES)
Specification
HEPATOCYTES
Treatment
20 ug/ml poly(I:C)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted Jul 09 2014

Western blot abreview for Anti-COX IV [20E8C12] antibody

 Average
Abreviews
abreview image
Application
Western blot
Loading amount
100000 cells
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (Jurkats)
Specification
Jurkats
Treatment
TCR activation with OKT3 and anti CD28
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Dr. Tejpal Gill

Verified customer

Submitted Jan 13 2014

Western blot abreview for Anti-COX IV [20E8C12] antibody

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (8-12)
Sample
Mouse Tissue lysate - whole (liver)
Specification
liver
Blocking step
No blocking step used for 30 minute(s) · Temperature: 30°C
Read More

Dr. Gillian Hunter

Verified customer

Submitted Aug 06 2013

Western blot abreview for Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (fly heart tube)
Specification
fly heart tube
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Read More

Abcam user community

Verified customer

Submitted Jul 02 2013

Western blot abreview for Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (h9c2)
Loading amount
30 µg
Specification
h9c2
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Read More

Abcam user community

Verified customer

Submitted Feb 18 2013

Western blot abreview for Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (hek293)
Loading amount
30 µg
Specification
hek293
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Read More

Abcam user community

Verified customer

Submitted Feb 18 2013

Question

Would like a good loading control for Zebrafish, B Tubulin etc

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Answer

We would recommend the use of one of the following products for a loading control with zebrafish samples: AB15224 anti alpha Tubulin (https://www.abcam.com/alpha-tubulin-antibody-ab15246.html). This product has been featured in the following publication for use in WB on zebrafish (PubMed ID: 22581286). AB6046 anti beta Tubulin. This product has an excellent Abreview provided by a customer using in WB on zebrafish. This review is located at the following web address: https://www.abcam.com/beta-Tubulin-antibody-Loading-Control-ab6046-reviews.html?intabreviewid=15893 AB14744 anti- COX IV antibody (https://www.abcam.com/COX-IV-antibody-20E8C12-Mitochondrial-Loading-Control-ab14744-references.html). Another guaranteed product for use as loading control this product has been used in publication (PubMed ID: 18765295).

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1-10 of 51 Abreviews or Q&A

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