Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7] (ab109864)

Overview

  • Product name
    Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7]
  • Description
    Mouse monoclonal [5F51BB5AG7] to Adenine Nucleotide Translocase Immunocapture
  • Host species
    Mouse
  • Tested applications
    Suitable for: IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Cow, Human
  • Immunogen

    Recombinant Human Adenine Nucleotide Translocase

  • Positive control
    • Bovine and Human heart mitochondria This antibody gave a positive result in IHC in the following FFPE tissue: Human normal heart muscle.
  • 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 ab109864 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
IP Use at 100 µg/mg of lysate.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Relevance
    ANT is a reversible transporter of ATP and ADP. During oxidative phosphorylation (OXPHOS) the protein exchanges ATP out for ADP in, however, in the absence of OXPHOS ANT works in reverse to maintain mitochondrial membrane potential. A subset of patients with heart related diseases have been shown to harbor mutations in the ANT gene or to develop an autoimmune reaction against the protein. There are a number of proteins found in mitochondria that, under circumstances favoring apoptosis, coalesce to form the so-called permeability transition pore. The proteins identified as a part of the PTP include cytosolic hexokinase, outer membrane porin (also called the voltage dependent anion channel or VDAC), adenylate cyclase in the intermembrane space, the adenine nucleotide transporter (ANT) and the peripheral benzodiazepine receptor protein of the inner membrane along with cyclophilin D of the matrix space. Hexokinase, porin and ANT all occur as isoforms and it remains to be determined whether there is specificity of these different forms for the PTP.
  • Cellular localization
    Mitochondrial

Images

  • IHC image of Adenine Nucleotide Translocase staining in Human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab109864, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunocapture of Adenine Nucleotide Translocase from Bovine heart mitochondria using ab109864 at a concentration of 100 µg/mg lysate.

References

This product has been referenced in:
  • Zhao X  et al. Bcl-xL mediates RIPK3-dependent necrosis in M. tuberculosis-infected macrophages. Mucosal Immunol 10:1553-1568 (2017). Read more (PubMed: 28401933) »
  • Mailloux RJ  et al. Hexokinase II acts through UCP3 to suppress mitochondrial reactive oxygen species production and maintain aerobic respiration. Biochem J 437:301-11 (2011). Read more (PubMed: 21554247) »
See all 5 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

I am very pleased to hear you would like to accept our offer and test ab110322 and ab109864 in IP on mouse tissue. This code will give you 1 free PRIMARY ANTIBODY for each code before the expiration date. To redeem this offer, please submit an Abreview for IP on mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. The antibody was raised against full length recombinant human ANT1, but the exact binding site is unknown. 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 unfortunately at this time Anti-Adenine Nucleotide Translocase 1 antibody [5F51BB5AG7] (ab110322) has not been tested for use in IP. However, this does not mean that it will not work, just that we have not tested this product in IP and to our knowledge, our customers have not either.If you would be interested in testing ab110322 in IP we have a testing discount that we would be able to offer you. This would involve you purchasing the antibody, testing it in IP with mouse samples, then sharing the results with us through an Abreview. Regardless of if the results are positive or negative, you would then be entitled to a free primary antibody of your choice from our catalogue.

More information on this offer can be found from the following link:www.abcam.com/collaborationdiscount

If you would be interested in this offer please do let me know as I will need to issue you with a discount code prior to purchase of ab110322.

In regards to ab109864, this productuses the anti-ANT monoclonal antibody ab110322, therefore the beads should be able to IP ANT from mouse samples.However this has also not been tested. If you would be interested in testing this on mouse please let me know. The same program outlined above would be applicable here.

If you should choose ab109864, because this product is a mouse monoclonal and your tissue samples will be mouse, I would recommend use of a light chain only secondary antibody for the western blots. These would allow you to be confident of your band of interested is not obstructed by the heavy chain. As ab109864 is kappa, the correct seconddaries would be ab99617 (https://www.abcam.com/Rat-monoclonal-187-1-Secondary-Antibody-to-Mouse-kappa-light-chain-HRP-ab99617.html) or ab99632 (https://www.abcam.com/Rat-monoclonal-H139-52-1-Secondary-Antibody-to-Mouse-kappa-light-chain-HRP-ab99632.html).

I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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Answer

Thanks for your phone call last week and I apologize for the extended delay getting back to you. There is an error on the datasheet, and the recommended antibody ratio of 100 ug per mg of sample is actually for a different Mitosciences product in which the antibody is already coated to agarose beads. For ab109864, you can follow the general Abcam IP protocol - https://www.abcam.com/index.html?pageconfig=resource&rid=11385 For the primary antibody, we recommend titrating the optimal antibody ratio between 1ug and 100ug antibody per mg of sample depending on the sample type.    This antibody can work in Western blot and is most optimal following this specific protocol- http://www.mitosciences.com/PDF/ant_western_blotting_protocol.pdf I hope this information is helpful. Please let me know if you have any questions or if there is anything else we can do for you.

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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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