Anti-Polyoma virus, Medium T antigen antibody [PyMT] (ab15085)

Reviews (1)Q&A (7)References (9)

Overview

  • Product name
    Anti-Polyoma virus, Medium T antigen antibody [PyMT]
  • Description
    Rat monoclonal [PyMT] to Polyoma virus, Medium T antigen
  • Host species
    Rat
  • Tested applications
    Suitable for: IP, ICC/IF, WBmore details
  • Immunogen

    Synthetic peptide (N terminal).

  • General notes


    Binds medium T antigen only, allows isolation of viral T antigens.

Properties

Applications

Our Abpromise guarantee covers the use of ab15085 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 an assay dependent dilution.
ICC/IF Use at an assay dependent dilution.
WB Use at an assay dependent concentration.

Target

  • Relevance
    Middle T antigen (MT) is a 421-amino-acid protein associated with membranes and underlying cytoskeletal elements, and is associated with a tyrosine-specific protein kinase activity. It is the principal oncoprotein of polyomavirus that is necessary and often sufficient for transformation in vitro. MT delivered as a transgene or a retrovirus can induce tumors in a wide variety of tissues. Polyomavirus (PyV) is a small, double-stranded, closed-circular-DNA virus with an approximately 5-kb genome divided into two roughly equal regions. The late transcripts produce the viral capsid proteins, whereas the early region encodes three so-called tumor (T) antigens that are important for both productive infection and transformation.
  • Cellular localization
    Cytoplasmic location in cells infected with virus.
  • Alternative names
    • Middle T antigen antibody
    • MT AG antibody
    • MT antibody
    • Polyoma virus middle t antigen antibody

References

This product has been referenced in:
  • Kim J  et al. Ablation of miR-10b Suppresses Oncogene-Induced Mammary Tumorigenesis and Metastasis and Reactivates Tumor-Suppressive Pathways. Cancer Res 76:6424-6435 (2016). ICC/IF . Read more (PubMed: 27569213) »
  • Roy A  et al. Targeting Serglycin Prevents Metastasis in Murine Mammary Carcinoma. PLoS One 11:e0156151 (2016). Read more (PubMed: 27223472) »
See all 9 Publications for this product

Publishing research using ab15085? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-8 of 8 Abreviews or Q&A

Question

Product code: 15085
Lot number: GR37142-3

Inquiry: I would like to have some suggestion regarding the use of the above mentioned Ab in immunofluorescence. We stain the lungs to look for metastasis and visualize it with Alexa568 secondary. But the PyMT Ab gives a lot of background and some non-specific signals. We have narrowed down the dilution to 1:500 for the best results as of now. But I would like to make sure that we do not get any non specific signals at all so that we can count the metastasis from the signals without doubt.

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results.

This information will also allow us to investigate this case internally and initiate additional testing where necessary.

I look forward to receiving your reply.

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Question

Phone call requesting information of how the Anti-Polyoma virus, Medium T antigen antibody [PyMT] (ab15085) has been used in ICC.

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Answer

I am sorry for the delay in getting back to you in regards to the protocol used with the Anti-Polyoma virus, Medium T antigen antibody [PyMT] (ab15085).
I can now provide some further guidance in how I would suggest using the antibody in IHC. From the information I have I would suggest initially trying the antibody at a dilution of 1/100. From this you should be able to ascertain if you can reduce the concentration further, or if you do not get any signal, you may have to decrease the dilution.
As we discussed over the phone, antigen retrieval may be required and this may need some optimisation to get the best results. We have quite a comprehensive guide which can give you a few tips on how this can be approached:
https://docs.abcam.com/pdf/misc/abcam-protocols-book-2010.pdf
If I can be of any further assistance, please do let me know. Until then, I wish you all the best with your research.

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Question

Phone call requesting information of how the Anti-Polyoma virus, Medium T antigen antibody [PyMT] (ab15085) has been used in ICC.

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Answer

Thank you for contacting us yesterday in regards to the Anti-Polyoma virus, Medium T antigen antibody [PyMT] (ab15085).
Unfortunately, as we discussed over the phone, this antibody has as yet not been used in IHC to our knowledge. This does not mean that it would not work, simply that we have not tried it ourselves and would not be able to guarantee its performance in this application. However, if you would like to purchase ab15085 and use it in IHC-P, we can offer you a testing discount. This offer means that if you purchase ab15085 as normal, test the antibody in IHC-P and let us know of the results through an Abreview (no matter whether positive or negative) you would be eligible for a free primary antibody of your choice from our catalogue (or the equivalent price of ab15085 off any product). More information on this offer can be found here:
www.abcam.com/collaborationdiscount
Please note that the antibody to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.
I have generated the testing discount code as follows:
Discount code: xxxxx
Expiration date: 16th of January 2013.
Please quote this code in your Abreview for ab15085 to activate it.
As discussed over the phone, I am seeking further information as to how the antibody was used in ICC and will forward this information to you once I have it.
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

Please see technical inquiry from our customer:

I’m interested in product ab15085 Anti-Polyoma virus, Medium T antigen antibody [PyMT].

I like to know whether this antibody will also be able to detect the Large and small T antigen? From my understanding, middle T antigen contains 421 amino acids and is divided into at least three domains, some of which are shared with the Large T antigen and small T antigen. Having said that, would using product ab15085 have the epitopes to detect Large, medium, and small T antigen ?

I want to apply this antibody in western blot to detect total PyMT in samples.

Looking forward for your response.

Kind regards,

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Answer

Thank you very much for your patience.
We can confirm that the antibody is specific to Medium T form only. We have no cross reactivity data regarding small and large T forms. The clone specific publication data show the antibody should not be cross reacting with the small and long forms.
I hope this information is nevertheless helpful to you . Should you have any question please do not hesitate to contact me.

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Question

Hi,
we are using antibody ab15085 for IF on formalin-fixed paraffin-embedded tissues, but we always get tons of autofluorescence. do you have good protocol for this?
btw, does this antibody can be used for IHC? or any other polyomavirious middle T antigen antibody can work for IHC?
thanks a lot.

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Answer

Thank you for contacting us.

I am afraid ab15085 has not been tested in IHC, and therefore we cannot guarantee it will work in this application. You may find useful our online protocols about IHC in order to optimise the staining:

https://www.abcam.com/index.html?pageconfig=popular_protocols

Unfortunately this is the only antibody available against the Medium T antigen from Polyoma virus.

Autofluorescence may be a consequence of the tissue itself, or even the fixative. You can try to optimise the results by reducing antibody concentration, washing the slides adequately, use serum from the species in which the secondary was raised to block unspecific bindings and depending on the tissue and the conditions used, blocking of endogenous enzymes may also be useful.

It is also crucial to use a positive control to assess the source of the problem

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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Western blot abreview for Anti-Polyoma virus, Medium T antigen antibody [PyMT]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Breast tumor)
Loading amount
50 µg
Specification
Breast tumor
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Jan 27 2011

Question

Lot # 145691 I think that the purchase order number is UR114911, if not then ill talk to the administrators The PO number is 346756

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Answer

Thank you for your email and patience. I presently have concerns about the suitability of this antibody in Western blotting and have removed the application from our online datasheet. We haven't received any other complaints, but the originator of this antibody has told me that it works "weakly" in WB and there is no additional information regarding this. I would be happy to offer you a credit note or refund, please let me know which you would prefer and I look forward to hearing from you. Have a great weekend.

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Question

Are there more details available about how this antibody was tested in histology?

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Answer

Thank you for your enquiry. More details about this antibody can be found in the following references: Dilworth SM & Griffin BE Monoclonal antibodies against polyoma virus tumor antigens. Proc Natl Acad Sci U S A 79:1059-63 (1982). PubMed: 6175960 Dilworth SM Protein kinase activities associated with distinct antigenic forms of polyoma virus middle T-antigen. EMBO J 1:1319-28 (1982). PubMed: 6327264

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