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

Submit a review Q&A (3)References (5)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7] (ab109864)
  • Immunoprecipitation - Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7] (ab109864)

Key features and details

  • Mouse monoclonal [5F51BB5AG7] to Adenine Nucleotide Translocase Immunocapture
  • Suitable for: IP, IHC-P
  • Reacts with: Cow, Human
  • Isotype: IgG1

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 full length protein. This information is proprietary to Abcam and/or its suppliers.

  • 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. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Product was previously marketed under the MitoSciences sub-brand.

Properties

Applications

The Abpromise guarantee

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.
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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7] (ab109864)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7] (ab109864)

    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.

  • Immunoprecipitation - Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7] (ab109864)
    Immunoprecipitation - Anti-Adenine Nucleotide Translocase Immunocapture antibody [5F51BB5AG7] (ab109864)
    Immunocapture of Adenine Nucleotide Translocase from Bovine heart mitochondria using ab109864 at a concentration of 100 µg/mg lysate.

References (5)

Publishing research using ab109864? Please let us know so that we can cite the reference in this datasheet.

ab109864 has been referenced in 5 publications.

  • Zhao X  et al. Bcl-xL mediates RIPK3-dependent necrosis in M. tuberculosis-infected macrophages. Mucosal Immunol 10:1553-1568 (2017). 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). PubMed: 21554247
  • Shulga N  et al. Sirtuin-3 deacetylation of cyclophilin D induces dissociation of hexokinase II from the mitochondria. J Cell Sci 123:894-902 (2010). PubMed: 20159966
  • Martin LJ  et al. The mitochondrial permeability transition pore in motor neurons: involvement in the pathobiology of ALS mice. Exp Neurol 218:333-46 (2009). PubMed: 19272377
  • Murray J  et al. Focused proteomics: towards a high throughput monoclonal antibody-based resolution of proteins for diagnosis of mitochondrial diseases. Biochim Biophys Acta 1659:206-11 (2004). PubMed: 15576053

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Question

thanks for the information. I would love to try the IP conditions and would love to test both antibodies ab110322 as well as ab109864 for IP purpose on mouse tissue. Do you know the peptide the Ab is directed against? I would like to test for interaction with another protein and would like to know where the peptide the ab is directed against is localized.

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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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Question

can this antibody also used for IP purpose? (Anti-Adenine Nucleotide Translocase 1 antibody [5F51BB5AG7] (ab110322)). or does the one recommended for IP purpose, recognize mouse? Against which peptide is this one directed at (MSA01c) thanks a lot,

Read More
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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Question

Is the recommended concentration for IP correct (100 ug per 1 mg of lysate)? I frequently use 1-5 ug of antibody per mg of mitochondrial lysate. Is the antibody suitable for Western blot?

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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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