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

Reviews (23)Q&A (24)References (223)

Key features and details

  • Mouse monoclonal [20E8C12] to COX IV
  • Suitable for: Flow Cyt, IHC-FoFr, WB, ICC/IF, IHC-Fr, IHC-P, IP
  • Reacts with: Mouse, Rat, Hamster, Cow, Human, Pig, Drosophila melanogaster, Zebrafish
  • Isotype: IgG2a

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)
    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

  • 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.

  • 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

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

ab14744 has been referenced in 223 publications.

  • Hara T  et al. Use of in-gel peroxidase assay for cytochrome c to visualize mitochondrial complexes III and IV. Biol Open 9:N/A (2020). PubMed: 31852667
  • Ma L  et al. Sennoside A restores colonic barrier function through protecting colon enterocytes from ROS-induced mitochondrial damage in diet-induced obese mice. Biochem Biophys Res Commun 526:519-524 (2020). PubMed: 32245617
  • Yang YD  et al. Nogo-B Receptor Directs Mitochondria-Associated Membranes to Regulate Vascular Smooth Muscle Cell Proliferation. Int J Mol Sci 20:N/A (2019). PubMed: 31083380
  • Ghosh A  et al. SCAN1-TDP1 trapping on mitochondrial DNA promotes mitochondrial dysfunction and mitophagy. Sci Adv 5:eaax9778 (2019). PubMed: 31723605
  • Yuan Z  et al. Koumine Promotes ROS Production to Suppress Hepatocellular Carcinoma Cell Proliferation Via NF-?B and ERK/p38 MAPK Signaling. Biomolecules 9:N/A (2019). PubMed: 31581704
View all Publications for this product

Customer reviews and Q&As

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1-10 of 47 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?

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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.

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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

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

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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Question

Hello again,

Thank you for all the help and advice that you and other members of your company have provided. I have yet another request. I am looking for a monoclonal Ab that would react with Human and mouse cell lines and would be specific for a nuclear membrane protein that is located on the outer membrane. As many as you can recommend would be great.

Thank you,

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Answer

Thank you very much for your reply.

The common nuclear outer membrane proteins are nesprins, NAV3, and OPRS1. The nesprins and OPRS1 can also be found in the inner membrane or other organelles, but the NAV3 seems to be more specific to the outer membrane. Unfortunately we do not have any monoclonal antibodies to this target, just polyclonals such as ab101792-

https://www.abcam.com/nav3-antibody-n-terminal-ab101792.html

We do have monoclonal antibodies to nesprins, such as ab125931-

https://www.abcam.com/nesprin-1-antibody-mannes1a7a12-ab125931.html

Would either of these work for you?

I am sorry that we don't have more options at this time, but please let me know if there is anything else that I can help find for you, and I'll be happy to help.

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Question

Hello,

I am still hunting for some decent antibodies that I can use to verify findings by Western. I am interested in polyclonal Abs that react with Human and Mouse cell lines against membrane bound proteins of Nuclei and Mitochondria . I want to make sure that the proteins used to generate the Abs are not soluble. I have not had any luck using the polyclonal Nesprin Ab from you guys. I get more of a ladder with HeLa lysate rather than a specific 112 kDa band. The same is true for the polyclonal TOM22 Ab that gives me a band at 60 kDa rather than the specific band at 18 kDa.

Can you please suggest some Nuc and Mito Abs that are targeted against membrane bound proteins? These would preferably be polyclonal and react with both human and mouse. And the application would be for western, where there is one specific band.

Thank you,

Read More
Answer

Thank you very much for your enquiry.

I am sorry to hear that the nesprin and TOMM22 antibodies are not working as they should be. Could you tell me the catalog numbers of each of these antibodies, and also a description of the protocol that you're using? We do fully guarantee all of our products to work as stated on the datasheets, so I'd be happy to send replacements if applicable. Could you also send your original order or PO number?

If you'd like to continue probing for nesprin and TOMM22, we do have a couple of antibodies that work in Western blot with human and mouse samples-

https://www.abcam.com/TOMM22-antibody-ab57523.html

https://www.abcam.com/Nesprin3-antibody-ab74261.html

We also have several other antibodies that are targeted against mitochondrial or nuclear membrane-bound proteins that work in Western blot with mouse and human samples:

Mitochondrial

Prohibition antibody: https://www.abcam.com/Prohibitin-antibody-Mitochondrial-Marker-ab28172.html

MTCO1 antibody: https://www.abcam.com/MTCO1-antibody-1D6E1A8-Mitochondrial-Marker-ab14705.html

Nuclear

Lamin B1 antibodies: https://www.abcam.com/Lamin-B1-antibody-Nuclear-Envelope-Marker-ab16048.html

https://www.abcam.com/Lamin-B1-antibody-119D5-F1-Nuclear-Envelope-Marker-ab8982.html

Would one of these work for you?

I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

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Question

Thank you so much for your previous help. I have another inquiry now. I’m trying to isolate nuclei from mammalian cells and would like any advice on an antibody suggestion for this and more so for identifying said pull down by western. I have tried Histone Abs and would like something that is a bit bigger than 17 kDa but also easily distinguished from antibody chains that would be present in the pull down sample. And secondly, a good antibody that is polyclonal for identifying the presence of mitochondria in the above pull down sample, with same criteria as the Nuc western detection antibody described above.

Thank you so much.

Read More
Answer

Thank you very much for contacting us with your questions.

Will you be trying to isolate intact nuclei from mammalian cells? I'm not sure if there is an antibody that would be good for this, butthecell fractionation kit ab109719 that I mentioned beforemight be suitable. I'll need to check with the lab whether the nucleiu will still be intact using this kit, if that is your end goal.

To check the efficiency of a nuclear fractionation, I would recommend probing for either lamin B1 (66 kDa) or TATA binding protein (38 kDa). Both of these proteins will run higher than the histones but should be far enough away from the IgG heavy and light chains around 25 and 50 kDa. For mitochondria, I'd recommend a VDCA1/porin antibody (31 kDa), though this may be too close to the 25 kDa light chain band. You could also probe for COXIV(16 kDa) but this protein may be smaller than what you'd like. What species will your samples be from? I can find specific antibodies that are known to react with this species.

I'm not sure if you would be interested inthis, but we do have secondary antibodies that only bind to native primary antibodies so that the denatured chains won't be visible at 25 and 50 kDa following an IP. For anti-mouse IgG, we have ab131368-

https://www.abcam.com/mouse-IgG-VeriBlot-for-IP-secondary-antibody-HRP-ab131368.html

For anti-rabbit IgG, we have ab131366-

https://www.abcam.com/rabbit-IgG-VeriBlot-for-IP-secondary-antibody-HRP-ab131366.html

I look forward to hearing from you. Please let me know if you have any further questions or if there is anything else that we can do for you.

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Question

Do you know which kind of protein (GAPDH, actin…)can be used as an internal-control of western blotting for liver tissue mitochondria?I look forward to your reply.

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Answer

Unfortunately, we do not have a special "rat liver mitochondria loading control"but we have one antibody which may interest you:

ab16056 Anti-COX IV antibody - Mitochondrial Loading Control

https://www.abcam.com/index.html?datasheet=16056 (or use the following: https://www.abcam.com/index.html?datasheet=16056).

This antibody has two very positive Abreviews in WB and rat, and has been demonstrated to work in liver, however, the latter wasin IHC and human. Nonetheless, I think this is the right one for you in this case.

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1-10 of 47 Abreviews or Q&A

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