Overview

  • Product name
    Anti-ATP5J antibody [10C6AC9]
    See all ATP5J primary antibodies
  • Description
    Mouse monoclonal [10C6AC9] to ATP5J
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant Human ATP5J protein (UniProt ID: P18859).

  • Positive control
    • WB: Isolated mitochondria from Human heart lysate. ICc/IF: Cultured Human embryonic lung-derived fibroblasts (strain MRC5) Flow Cyt: HL-60 cells
  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    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 ab110279 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
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 13 kDa.
Flow Cyt Use a concentration of 1 µg/ml.

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

 

ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    Mitochondrial membrane ATP synthase (F(1)F(0) ATP synthase or Complex V) produces ATP from ADP in the presence of a proton gradient across the membrane which is generated by electron transport complexes of the respiratory chain. F-type ATPases consist of two structural domains, F(1) - containing the extramembraneous catalytic core and F(0) - containing the membrane proton channel, linked together by a central stalk and a peripheral stalk. During catalysis, ATP synthesis in the catalytic domain of F(1) is coupled via a rotary mechanism of the central stalk subunits to proton translocation. Part of the complex F(0) domain and the peripheric stalk, which acts as a stator to hold the catalytic alpha(3)beta(3) subcomplex and subunit a/ATP6 static relative to the rotary elements. Also involved in the restoration of oligomycin-sensitive ATPase activity to depleted F1-F0 complexes.
  • Sequence similarities
    Belongs to the eukaryotic ATPase subunit F6 family.
  • Cellular localization
    Mitochondrion. Mitochondrion inner membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATP synthase, H+ transporting, mitochondrial F0 complex, subunit F6 antibody
    • ATP synthase-coupling factor 6, mitochondrial antibody
    • ATP synthase-coupling factor 6, mitochondrial antibody
    • ATP5 antibody
    • ATP5A antibody
    • ATP5J antibody
    • ATP5J_HUMAN antibody
    • ATPase subunit F6 antibody
    • ATPM antibody
    • CF6 antibody
    • F6 antibody
    see all

Images

  • Anti-ATP5J antibody [10C6AC9] (ab110279) at 1 µg/ml + Isolated mitochondria from Human heart lysate at 5 µg

    Predicted band size: 13 kDa

  • Mitochondrial localization of ATP synthase visualized by immunocytochemistry using ab110279.
    Cultured Human embryonic lung-derived fibroblasts (strain MRC5) were fixed, permeabilized and then labeled with ab110279 (1 µg/ml) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG(H+L) secondary antibody (2 µg/ml).
  • HL-60 cells were stained with 1 µg/mL ab110279 (blue) or an equal amount of an isotype control antibody (red) and analyzed by Flow Cytometry.
  • All lanes : Anti-ATP5J antibody [10C6AC9] (ab110279)

    Lane 1 : human heart homogenate at 20 µg
    Lane 2 : HepG2 cell lysate at 20 µg
    Lane 3 : human liver mitochondrial extraction at 10 µg
    Lane 4 : bovine heart mitochondrial extraction at 10 µg
    Lane 5 : human heart mitochondrial extraction at 10 µg
    Lane 6 : human liver homogenate at 20 µg

    Predicted band size: 13 kDa



    Although the ATP5J gene has alternatively spliced transcript variants which encodes different isoforms, the MW of the isoforms should within 10~12kDa. The high MW band detected in liver tissues as well as in bovine heart may be a structural complex which involves ATP5J.

References

This product has been referenced in:
  • Pecina P  et al. Role of the mitochondrial ATP synthase central stalk subunits ? and d in the activity and assembly of the mammalian enzyme. Biochim Biophys Acta 1859:374-381 (2018). Read more (PubMed: 29499186) »
  • Akbergenov R  et al. Mutant MRPS5 affects mitoribosomal accuracy and confers stress-related behavioral alterations. EMBO Rep N/A:N/A (2018). WB ; Human . Read more (PubMed: 30237157) »
See all 3 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for your email, I am sorry for the delay in getting back to you.

I have a couple of antibodies your customer may be interested in:

Anti-Frataxin antibody [18A5DB1] (ab110328)

and

Anti-ATP5J antibody [10C6AC9] (ab110279)

These have been tested in house with mouse tissue and found not to cross react. They are however able to detect the human mitochondrial proteins. Unfortunately we are not able to guarantee this as the homology of the immunogens used to raise the proteins is 73 to 80% respectively. However, if your customer would like to try it I can offer them the Abtrial testing discount:

https://www.abcam.com/abtrial

I would just like to point out if they are intending to use the antibodies in IHC both antibodies have been raised in mouse and they would need to take into consideration the possibility of higher background caused by mouse-on-mouse staining. We have kits which can help in this, as well as protocol advice:

Mouse on Mouse Polymer IHC Kit (ab127055)

Mouse on mouse staining procedure:

https://www.abcam.com/index.html?pageconfig=resource&rid=11465

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

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Answer

Merci beaucoup pour votre patience. Le laboratoire avait retesté cet anticorps sur différents tissus. Veuillez trouver les résultats dans le pièce jointe. Effectivement nous sommes très confiantes que cet anticorps reconnaît le ATP5J, car il était développée contre cette protéine. Nous sommes par contre aussi d'accord que la protéine devrait sortir à 12kD. Le fait qu'elle apparaît vers les 25kD dans certains tissus, nécessite une investigation plus poussé. C'est peut-être des complexes de protéines.   Effectivement aussi le ab65841 donne une bande aux alentours de 20kD sur des extraits du foie. Est-ce que vous pouvez me dire ce que vous en pensez? Est-ce que vous auriez la possibilité de vérifier cet anticorps sur des extraits de coeur? La légende de l'images est: HHH (human heart homogenate), HepG2(HepG2 cell lysate), HLM (human liver mitochondrial extraction), BHM (bovine heart mitochondrial extraction), HHM (human heart mitochondrial extraction), and HLH (human liver homogenate). Je me réjouis de recevoir de vos nouvelles.

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Answer

Je suis désolée d'apprendre que ce tube de l'ab110279 ne donne pas de bons résultats en Western blot.  Veuillez trouver ci-joint le questionnaire discuté au téléphone. Veuillez le nous retourner avec une image du résultat. Comme discuté aussi, je vais ensuite investiguer avec les producteurs de cet anticorps ce problème. Si nous trouvons pas une solution et concluons que notre produit est fautif , nous serons ravis de mettre en place l'envoi d'un tube de remplacement gratuit ou d'un remboursement. Je vous remercie par avance de nous envoyer votre image et protocole.  

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Answer

DISCOUNT CODE: XXXXXXXXX Expiration date: 15-01-2012 I am very pleased to hear you would like to accept our offer and test ab110279 in Mouse. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for Mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us. We have not tested this antibody with mouse lysates yet so we unfortunately are unable to share any tested data. The mouse and human protein have 80% homology so it is more likely that this antibody could detect the mouse protein also. As mouse is an untested species so this antibody is eligible for our Abreview testing discount programme which states, in case customers would like to use antibodies in untested species, they can receive a free primary antibody in exchange of an Abreview. Please be advised that the free of charge product will be your next purchase; the current purchase has to be at full price. Let me know if you are interested in using this antibody with mouse lysates I will then be creating a discount code for you which will give you your next primary ab free of charge. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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