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
  • 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

  • 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

  • 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

  • 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.

  • 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

  • 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. 

  • 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 187 Publications for this product

Customer reviews and Q&As

21-30 of 51 Abreviews or Q&A

Answer

Thank you for your enquiry. It has taken some time to confirm all the details for you, and I appreciate your patience.

I am sorry to confirm that MS602 has been discontinued and we do not have anymore in stock.

Although that product is not available any longer, there is ab110411, Total OXPHOS Human WB Antibody Cocktail and ab110416, Pyruvate dehydrogenase (PDH) WB Antibody Cocktail, however as the name suggests they have only been tested in WB, not IHC-P.

If you would like to considering trying to recreate the original antibody cocktail,I have listed the individual components are below.

Complex I 17G3D9E12 ab110243
https://www.abcam.com/index.html?datasheet=110243 (or use the following: https://www.abcam.com/index.html?datasheet=110243).

Complex II ab14714
https://www.abcam.com/index.html?datasheet=14714 (or use the following: https://www.abcam.com/index.html?datasheet=14714).

Complex III 13G12AF12BB11 ab14745
https://www.abcam.com/index.html?datasheet=14745 (or use the following: https://www.abcam.com/index.html?datasheet=14745).

Complex IV 1D6E1A8 ab14705
https://www.abcam.com/index.html?datasheet=14705 (or use the following: https://www.abcam.com/index.html?datasheet=14705).

Complex V 4C11C10D12 ab110276
https://www.abcam.com/index.html?datasheet=110276 (or use the following: https://www.abcam.com/index.html?datasheet=110276).

PDH 9H9AF5 ab110330
https://www.abcam.com/index.html?datasheet=110330 (or use the following: https://www.abcam.com/index.html?datasheet=110330).

As there are several components, we would be able to offer you a discount if you wish to purchase all of these products together. I am forwarding this to our bulk orders team who will be able to provide a quote for you.

Otherwise, I am sorry there is nothing else to suggest that would be a suitable replacement on this occasion.

I hope this will be helpful. If you have any further questions, please do not hesitate to contact me.

Read More

Answer

Thank you for your reply. It is possible that the epitope recognized by the monoclonal antibody ab14744 may be sensitive to the extraction protocol you are using. Switching to a polyclonal antibody such as ab16056 may give better results by binding to multiple epitopes on COX IV. I would be happy to send you a vial of ab16056 as a free of charge replacement if you are interested in trying it. Please let me know your original order number and how you would like to proceed and I will be happy to assist you further.

Read More

Answer

Thank you for your reply.  Could you please provide some details about your extraction protocol?  Diffuse bands can sometimes be caused when the protein is altered by exposure to different pH levels.  Have you tried this experiment using mitochondrial extracts that were obtained during a different isolation?  If you are unsatisfied with your results, please let me know and I would be happy to offer you a free of charge replacement with one of our other COX IV antibodies or a credit or refund.  Please let me know how you would like to proceed and I will be happy to assist you further. 

Read More

Answer

Thank you for contacting us. Abcam takes product quality seriously and we recently undertook an in-depth study examining storage conditions and antibody performance. After over 13,000 individual ELISA experiments, we have determined that storing our antibodies at various temperatures of up to 45°C for 1 week does not impact their activity. As a precautionary measure only, we ship our antibodies in packaging with ice packs to provide extra temperature stability in transit. The ice pack may be thawed or even at room temperature when you receive it; please be assured that this is normal and that your product is safe to use. Once you have received the vial, please follow the long-term storage instructions on the datasheet. Our Abpromise to you is that, should our product not work as stated on the datasheet, we will resolve the issue to your satisfaction, either with helpful advice to optimize your experiment or a replacement or refund of the product. Please contact our knowledgeable Scientific Support staff should you experience any issues while using our products. Contact information can be found at: https://www.abcam.com/index.html?pageconfig=contactus I hope this information helps, but please don't hesitate to contact me for additional information or assistance.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (colonic epithelial cells)
Specification
colonic epithelial cells
Fixative
Paraformaldehyde
Permeabilization
Yes - Saponin 0.5%
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 07 2011

Question
Answer

Thank you for contacting us. I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways: (1) Redeemed against the original invoice if this hasn't already been paid. (2) Held on the account for use against a future order. (3) A full refund can be offered where no other invoices are outstanding. Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge. To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website. The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice  

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Question

I recently purchased two antibodies from you and was very disappointed with how they performed. It was mouse anti CoxIV (ab14744) and mouse anti PurA (ab77734) antibodies. They were intended to be used for ICC/IF, but both gave very poor staining.   I have tried the mouse anti CoxIV antibody (ab14744) in both mouse and in human cells. The cells were fixed in 4% formaldehyde and I titrated the antibody dilution between 1:100 and 1:5000, and compared with your rabbit polyclonal to CoxIV antibody (ab16056). ab14744 gave virtually no staining while ab16056 gave very nice mitochondrial staining at 1:1000 dilution. It clearly states in the datasheet that ab14744 should work in ICC/IF and be reactive in both human and mouse. This antibody was mainly intended to be used as a control when running in situ proximity ligation assay (the Duolink kit from Olink), which I mainly use on mES cells. Do you have any other mouse antibodies against any mitochondrial protein that is guaranteed to work in mouse cells for ICC/IF following formaldehyde fixation? Due to the poor performance of the above mentioned antibody I expect a steep discount on any such antibody.   My experience with the mouse anti PurA (ab77734) antibody is similar, but I only bought this antibody because I received a “Telomere antibodies” newsletter from you saying that it had been tested successfully in mouse on ICC/IF. In my hands however it gives no staining at all above background in mES cells, where on the contrary your rabbit anti PurA antibody (ab79936) gives a very strong nuclear staining mainly near the nuclear envelope.   I have been very pleased with your products and service in the past, but was very disappointed with the above mentioned products and thus hope to be fairly compensated.    

Read More
Answer

Thank you for contacting Abcam. I am sorry that you have been experiencing difficulties with our two antibodies, ab14744 and ab77734.  We have received positive feedback for both of these antibodies in ICC and so I am not sure why they have not worked well in your experiments.  I have checked our catalog, and unfortunately these are the only two mouse monoclonal antibodies against these targets.  I am happy to offer replacements, free of charge with a new vial of these antibodies or provide a credit for the products.  Please let me know which you would prefer. I hope this information is helpful and I do apologize for the inconvenience this issue has caused.  Please do not hesitate to contact us if you have any additional questions.  I look forward to your reply so that I may assist you further.

Read More

Question
Answer

Thank you for your enquiry. This antibody reacts well will COX-IV in all tissues tested, which is consistent with reactivity to isoform 1 which has an ubiquitous tissue expression pattern. In contrast, isoform 2 is expressed at high levels in lung. However, we have not ruled out cross-reactivity to isoform 2 as we do not have the pure protein available to do a definitive test. I hope this information will still be helpful. If there is anything else that I can help you with please do not hesitate to contact me.

Read More
Application
Western blot
Sample
Human Cell lysate - other (isolated mitochondria from HaCaT cells)
Loading amount
20 µg
Specification
isolated mitochondria from HaCaT cells
Treatment
50mJ/cm2 UV treatment
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 14 2011

Application
Western blot
Sample
Mouse Cell lysate - other (N2a)
Loading amount
5 µg
Specification
N2a
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 07 2011

21-30 of 51 Abreviews or Q&A

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