Overview

  • Product name
  • Description
    Rabbit polyclonal to VAP1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide derived from within residues 150 - 250 of Rat VAP1.

    Read Abcam's proprietary immunogen policy (Peptide available as ab42884.)

  • Positive control
    • This antibody gave a positive signal in the following Tissue Lysates: Lung (Rat), Lung (Mouse) and Lung (Human). This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung.

Properties

Applications

Our Abpromise guarantee covers the use of ab42885 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa).

Target

  • Function
    Cell adhesion protein that participates in lymphocyte recirculation by mediating the binding of lymphocytes to peripheral lymph node vascular endothelial cells in an L-selectin-independent fashion. Has a monoamine oxidase activity. May play a role in adipogenesis.
  • Tissue specificity
    Strongly expressed on the high endothelial venules of peripheral lymph nodes and on hepatic endothelia. Also highly expressed in appendix, lung and small intestine. Expressed also in adipose tissue, in bone marrow, colon, heart, kidney, ovary, pancreas, placenta, prostate, skeletal muscle, spleen and testis.
  • Sequence similarities
    Belongs to the copper/topaquinone oxidase family.
  • Post-translational
    modifications
    Topaquinone (TPQ) is generated by copper-dependent autoxidation of a specific tyrosyl residue.
    N- and O-glycosylated.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Amine oxidase, copper containing 3 (vascular adhesion protein 1) antibody
    • AOC3 antibody
    • AOC3_HUMAN antibody
    • Copper amine oxidase antibody
    • HPAO antibody
    • Membrane copper amine oxidase antibody
    • Membrane primary amine oxidase antibody
    • Semicarbazide sensitive amine oxidase antibody
    • Semicarbazide-sensitive amine oxidase antibody
    • SSAO antibody
    • VAP-1 antibody
    • Vascular adhesion protein 1 antibody
    • VP97 antibody
    see all

Images

  • IHC image of VAP1 staining in Human normal lung 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 ab42885, 1µg/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.

  • All lanes : Anti-VAP1 antibody (ab42885) at 1 µg/ml

    Lane 1 : Lung (Rat) Tissue Lysate
    Lane 2 : Lung (Mouse) Whole Cell Lysate - normal tissue
    Lane 3 : Lung (Human) Tissue Lysate - Fetal Normal Tissue

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 85 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 110 kDa. We are unsure as to the identity of these extra bands.



    VAP1 has a predicted molecular weight of 85 kDa; however it has a number of potential glycosylation sites which may affect the migration of the protein (SwissProt data). The observed 100kDa band is consistent with other commercially avaliable antibodies.

References

ab42885 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Answer

Thanks for your reply.

ab81673 has been tested for use in WB and IP so the experiment you propose should be fine. An alternative is to use one VAP1 antibody for IP and second VAP1 antibody for WB.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab48281.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Thanks for your reply.

Perhaps it would be more efficient to discuss this issue on the phone. Please feel free to call me. Alternatively, if you provide a phone number I'm happy to give you a call.

I look forward to speaking with you. I'm sure we will quickly come to an agreeable solution.

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Question


I attached our previous run of the lysate you sent us next to one we had. The one we’ve been using still produces a 75 kDa band, rather than the 85-100 kDa band that your website lists as VAP-1. As you can see, the lysate you sent us produces a much lower band under the same conditions.

It’s possible that this 75 kDa band is VAP-1, and for whatever reason we’re getting a slightly lower band than we should. We would believe this is the case if we could corroborate it with a positive control, but the lysate you sent us didn’t work as planned, either with the original antibody we bought or with the new antibody you gave us. It produced a much lower, 37 kDa or so band.

Abcam has sent us a free antibody and a free lysate, which we really appreciate, but they just have not worked in our hands the way your website guarantees they should. I’m not sure if this is a problem with the antibody itself, the lysate we had, or with our conditions, but it isn’t working the way we need it to. We think the lysate you gave us may be the issue because its results are very different from anything else we've run with either antibody. We’re currently using reducing conditions, so if there are other suggestions you may have that might help, that would be great.

The other antibody we have for VAP-1 is ab81673, which we haven’t used in western blotting because it isn’t guaranteed for western blot. This one does work fine for us for other uses.

We’d really like a different lysate to use as a positive control, because our other loading controls don’t produce a band in your recommended range.

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Answer

Thanks for your email.

Since you have ab81673 I would recommend using it in a western blot with the 2 lysates you have been testing. It is quite likely this antibody will work in western blotting as it has already been shown to work in a diverse application range.

Thanks for all your troubleshooting efforts. I look forward to hearing how this next western blot goes.

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Answer

Thanks for your email.

The data you are seeing do appear unexpected.

Reducing conditions are used during western blotting.

Before I believe a loading control western blot was producing expected banding pattern so I don't think this is an issue with the lysates. More likely it appears to be an issue with the antibody.

Have you used any other anti-VAP-1/AOC3 antibodies to test for reactivity via western blotting?

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Answer

Thanks for your reply.

While I'm happy to provide another vial of ab29297 or perhaps a different cell extract if you like, it appears that your in-house lung extract is working quite well. Perhaps it would be most efficient to continue using that lung lysate.

What do you think?

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Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1116832.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

**I will look into the availability of additional data using mouse whole lung lysate and follow up as soon as possible.

I wish you the best of luck with your research.

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Answer

Thanks for your email.

I am happy to send another VAP1 antibody. Please let me know which anti-VAP1 antibody you would like as a replacement, as well as the order reference number or PO number used to purchase the initial vial of ab42885.

Thanks in advance for the additional information!

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Answer

Thank you for that information.

You should receive the vial of lysate to use as a positive control tomorrow. If after you receive the lysate, you are still not seeing the correct size band, then please let me know and I will be happy to look into replacing the antibody itself for you.

In the meantime, if there is anything else I can help you with, please let me know.

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Answer

Thank you for calling Abcam earlier today.

I have been trying to see if there is a reason that you are getting a 75kDa band, instead of a band at 85kDa or 100kDa. I will be honest and say that I am unsure as to why that is occurring, but I was thinking that I could send a mouse lung lysate for you to use as a positive control:

https://www.abcam.com/Lung-Mouse-Whole-Cell-Lysate-normal-tissue-ab29297.html



If that would be ok with you, then please let me know and ill have it sent to you. Could you also please include your Abcam order number for ab42885 aswell.

I look forward to your reply and helping resolve this issue.

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