Anti-Adenine Nucleotide Translocase 1/ANT 1 antibody [5F51BB5AG7] (ab110322)

Overview

  • Product name

    Anti-Adenine Nucleotide Translocase 1/ANT 1 antibody [5F51BB5AG7]
    See all Adenine Nucleotide Translocase 1/ANT 1 primary antibodies
  • Description

    Mouse monoclonal [5F51BB5AG7] to Adenine Nucleotide Translocase 1/ANT 1
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human, Caenorhabditis elegans
  • Immunogen

    Recombinant full length protein corresponding to Human Adenine Nucleotide Translocase 1/ANT 1.

  • Positive control

    • Human, Bovine, Rat and Mouse isolated mitochondria; Cultured Human fibroblasts.
  • General notes

    This antibody comes with an Incubation Solution which should be used as the diluent for Western blot (the antibody should not be diluted in milk for incubations). The lab has found that diluting the antibody in this solution gives optimal results. Dilute the antibody to 1 ug/mL in neat Incubation Solution before use.

    The following is a quick reference procedure guide for use specifically with ab110322.

    1. Solubilize samples in SDS-PAGE sample buffer.

    2. Heat sample to 37°C for 30 minutes.

    3. Separate proteins on Tris-glycine 10-22% gradient gel 150 V ~ 2 hrs.

    4. Soak gel in 50 mL CAPS buffer for 30 minutes.

    5. Transfer to 0.45 µm PVDF membrane in CAPS buffer for 2 hrs at 0.15 A CAPS buffer : 10 mM 3-[cyclohexylamino]-1-propane sulfonic acid, pH 11 w/NaOH, 10% methanol ice cold throughout.

    6. Block membrane for 3 hrs - overnight in 5% milk/PBS 4°C.

    7. Wash blot 3 times for 5 mins PBS/0.05% Tween-20 with agitation. Followed by a final rinse in PBS.

    8. Antibody diluted to concentration specified in supplied incubation solution for 2 hrs.

    9. Wash blot 3 times in PBS/0.05% Tween-20 for 5 mins.

    10. Incubate blot with secondary anti-mouse antibody diluted according to manufacturer’s instructions in 1% milk/PBS for 2 hrs.

    11. Wash blot 3 times in PBS/0.05% Tween-20 for 5 mins. Rinse blot in PBS.

    12. Develop Blot: For alkaline phosphatase we recommend BCIP/NBT. For horseradish peroxidase we recommend ECL+.

    Previously labelled as Adenine Nucleotide Translocase 1. 

Properties

Applications

Our Abpromise guarantee covers the use of ab110322 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: 33 kDa.
ICC/IF Use a concentration of 10 µg/ml. (heat-induced antigen-retrieval improves signal)

Target

  • Function

    Catalyzes the exchange of ADP and ATP across the mitochondrial inner membrane.
  • Involvement in disease

    Defects in SLC25A4 are a cause of progressive external ophthalmoplegia with mitochondrial DNA deletions autosomal dominant type 2 (PEOA2) [MIM:609283]. Progressive external ophthalmoplegia is characterized by progressive weakness of ocular muscles and levator muscle of the upper eyelid. In a minority of cases, it is associated with skeletal myopathy, which predominantly involves axial or proximal muscles and which causes abnormal fatigability and even permanent muscle weakness. Ragged-red fibers and atrophy are found on muscle biopsy. A large proportion of chronic ophthalmoplegias are associated with other symptoms, leading to a multisystemic pattern of this disease. Additional symptoms are variable, and may include cataracts, hearing loss, sensory axonal neuropathy, ataxia, depression, hypogonadism, and parkinsonism.
  • Sequence similarities

    Belongs to the mitochondrial carrier family.
    Contains 3 Solcar repeats.
  • Cellular localization

    Mitochondrion inner membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • AAC1 antibody
    • Adenine Nucleotide Translocase 1 antibody
    • Adenine nucleotide translocator 1 (skeletal muscle) antibody
    • Adenine nucleotide translocator 1 antibody
    • ADP antibody
    • ADP ATP carrier protein 1 antibody
    • ADP ATP carrier protein heart/skeletal muscle isoform T1 antibody
    • ADP/ATP translocase 1 antibody
    • ADT1_HUMAN antibody
    • ANT 1 antibody
    • ANT antibody
    • ANT1 antibody
    • ATP carrier protein 1 antibody
    • ATP carrier protein antibody
    • heart/skeletal muscle isoform T1 antibody
    • PEO2 antibody
    • PEO3 antibody
    • SLC25A4 antibody
    • Solute carrier family 25 member 4 antibody
    • T1 antibody
    see all

Images

  • ab110322 at 10µg/ml staining Adenine Nucleotide Translocase 1/ANT 1 in cultured Human fibroblasts (fixed and permeabilized) by Immunocytochemistry, followed by a fluorescent goat-anti-mouse IgG.

  • All lanes : Anti-Adenine Nucleotide Translocase 1/ANT 1 antibody [5F51BB5AG7] (ab110322) at 1 µg/ml

    Lane 1 : Human heart mitochondria isolate at 5 µg
    Lane 2 : Bovine heart mitochondria isolate at 1 µg
    Lane 3 : Rat heart mitochondria isolate at 10 µg
    Lane 4 : Mouse heart mitochondria isolate at 10 µg

    Predicted band size: 33 kDa



    Extra bands in the mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.

References

This product has been referenced in:

  • Petrick HL & Holloway GP High intensity exercise inhibits carnitine palmitoyltransferase-I sensitivity to L-carnitine. Biochem J N/A:N/A (2019). Read more (PubMed: 30635360) »
  • Zhou H  et al. Inhibitory effect of melatonin on necroptosis via repressing the Ripk3-PGAM5-CypD-mPTP pathway attenuates cardiac microvascular ischemia-reperfusion injury. J Pineal Res N/A:e12503 (2018). Read more (PubMed: 29770487) »
See all 37 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (quadriceps muscle)
Total protein in input
100 µg
Immuno-precipitation step
Protein G
Specification
quadriceps muscle

Abcam user community

Verified customer

Submitted Jan 08 2018

Answer



I can confirm that the incubation solution does not contain any blocking reagent and should only be used to dilute the primary antibody ab110322. (1 ug/mL in neat solution.)

The following is a quick reference procedure guide for use specifically with ab110322.

1. Solubilize samples in SDS-PAGE sample buffer.

2. Heat sample to 37°C for 30 minutes.

3. Separate proteins on Tris-glycine 10-22% gradient gel 150 V ˜ 2 hrs.

4. Soak gel in 50 mL CAPS buffer for 30 minutes.

5. Transfer to 0.45 µm PVDF membrane in CAPS buffer for 2 hrs at 0.15 A CAPS buffer : 10 mM 3-[cyclohexylamino]-1-propane sulfonic acid, pH 11 w/NaOH, 10% methanol ice cold throughout.

6. Block membrane for 3 hrs - overnight in 5% milk/PBS 4°C.

7. Wash blot 3 times for 5 mins PBS/0.05% Tween-20 with agitation. Followed by a final rinse in PBS.

8. Antibody diluted to concentration specified in supplied incubation solution for 2 hrs. (1 ug/mL in neat incubation solution.)

9. Wash blot 3 times in PBS/0.05% Tween-20 for 5 mins.

10. Incubate blot with secondary anti-mouse antibody diluted according to manufacturer’s instructions in 1% milk/PBS for 2 hrs.

11. Wash blot 3 times in PBS/0.05% Tween-20 for 5 mins. Rinse blot in PBS.

12. Develop Blot: For alkaline phosphatase we recommend BCIP/NBT. For horseradish peroxidase we recommend ECL+.

Read More

Answer

Thank you for contacting us.

During development of this product many different optimization step where attempted. It was found that the protocol detailed on the datasheet gave the best results with this product. We highly recommend using this protocol when using ab110322 and cannot guarantee the effectiveness of this product if another protocol is used.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question
Answer

Thank you very much for your interest in ab110322 (MSA02) and sorry about the confusion with the catalog numbers.

To our knowledge, the Anti-Adenine Nucleotide Translocase 1 antibodyab110322 (or MSA02)has not been tested in immunoprecipitation. Therefore, I can offer a discount off a future purchase if you buy ab110322 now, test it in immunoprecipitation and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab110322 in immunoprecipitation. I will then send a discount code. This code must be issued before purchasing ab110322 so please wait for my reply before ordering.

2. Purchase ab110322 either by phone, fax, or online (https://www.abcam.com).

3. Test it in immunoprecipitation.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab110322 turns out to be unsuitable for immunoprecipitation, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: https://www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us and your interest in our products. There are a few differentpositive controlsI can suggest for using withthese antibodies.

I can sumarise as follows:

The anti-eNos(phosphoS847) antibody, ab16650,has been used with several different tissue lysates and shown a good signal. We do not have any proteins in our catalogue which would be suitable as a control for the phosphorylated protein and the nNOS (neural protein)https://www.abcam.com/nNOS-neuronal-protein-ab59110.htmlwould not be suitable as it is not phosphorylated. We have seen good signal with this antibodyin mouse spinal chord, cortex and forebrain. Similarly, the equivalent tissues in rat would also be expected to react well. This is a very large protein ˜160 kDa. Optimisation of the transfer conditions may therefore have to be performed to achieve the best results. Some tips in how to do this can be accessed from the following link:

https://www.abcam.com/index.html?pageconfig=resource&rid=13111

Similarly with the anti-Adenine Nucleotide Translocase 1 antibody [5F51BB5AG7] (ab110322), we have no suitable recombinant protein available from our catalogue suitable as a positive control. However, a good signal has been observed with both rat and mouse heart tissue. Please note that there are specific protocol recommendations with this antibody on the datasheet to achieve the best results with the antibody.

With the anti-eNOS antibody (ab66127) we do have a recombinant protein which maybe suitable as a positive control. This antibody is raised against a synthetic peptide taken froman internal internal sequence of human eNOS and the recombinant protein we have available includes the residues 61-160. In order to check if thisprotein is suitable as a positive controlI am contacting the source of this antibody to confirm this. I will let you know once I have this confirmation. Otherwise, this antibody has been used with mouse lung as a positive control.

I would just like to add that all the antibodies you have chosen have been tested with both mouse and rat samples and in Western blotting and would therefore be covered by the Abpromise for the experiments you are hoping to achieve. So if you do have any problems with any of the antibodies please do let us know. More information on the Abpromise can be found from the following link:

https://www.abcam.com/abpromise

I will get back to you with more information regarding the eNOS protein as soon as I can. I am sorry for the delay and any inconvenience this may cause you.

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Question
Answer

Thank you for contacting us.

I can confirm that the solution is an antibody diluent. The lab has found that diluting the antibody in this solution gives an optimal signal, and they recommend diluting the antibody to 1 ug/mL in neat incubation solution.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for your phone call.
I have attached the specific WB protocol for ab110322 to this email and am working to have it added to our datasheet. Our MitoSciences team does not have a specific protocol for ab110413, but I have attached their general WB protocol as well.
The incubation solution for ab110322 does not contain any protein blocking agents. I hope this helps, please let me know if you need any additional information or assistance.

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Answer

Thank you for your call today, and I apologize that we did not already have this information in our notes.
I've gotten in touch with the lab, and this is intended to be an antibody diluent for Western blot. The lab has found that diluting the antibody inthis solution gives an optimal signal, andthey recommend diluting the antibody to 1 ug/mL in neat incubation solution.
I hope that this information will be useful, but please let me know if you have any further questions or if there is anything else that we can do for you, and I'll be happy to help.

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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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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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1-10 of 14 Abreviews or Q&A

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