Anti-Wilms Tumor Protein antibody (ab15249)

Reviews (2)Q&A (15)References (19)

Overview

  • Product name
    Anti-Wilms Tumor Protein antibody
    See all Wilms Tumor Protein primary antibodies
  • Description
    Rabbit polyclonal to Wilms Tumor Protein
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide within Human Wilms Tumor Protein aa 400 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P19544

Properties

Applications

Our Abpromise guarantee covers the use of ab15249 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-Fr Use at an assay dependent concentration. PubMed: 22586490
IHC-P 1/100.

Target

  • Function
    Transcription factor that plays an important role in cellular development and cell survival. Regulates the expression of numerous target genes, including EPO. Plays an essential role for development of the urogenital system. Recognizes and binds to the DNA sequence 5'-CGCCCCCGC-3'. It has a tumor suppressor as well as an oncogenic role in tumor formation. Function may be isoform-specific: isoforms lacking the KTS motif may act as transcription factors. Isoforms containing the KTS motif may bind mRNA and play a role in mRNA metabolism or splicing. Isoform 1 has lower affinity for DNA, and can bind RNA.
  • Tissue specificity
    Expressed in the kidney and a subset of hematopoietic cells.
  • Involvement in disease
    Defects in WT1 are the cause of Frasier syndrome (FS) [MIM:136680]. FS is characterized by a slowly progressing nephropathy leading to renal failure in adolescence or early adulthood, male pseudohermaphroditism, and no Wilms tumor. As for histological findings of the kidneys, focal glomerular sclerosis is often observed. There is phenotypic overlap with Denys-Drash syndrome. Inheritance is autosomal dominant.
    Defects in WT1 are the cause of Wilms tumor 1 (WT1) [MIM:194070]. WT is an embryonal malignancy of the kidney that affects approximately 1 in 10'000 infants and young children. It occurs both in sporadic and hereditary forms.
    Defects in WT1 are the cause of Denys-Drash syndrome (DDS) [MIM:194080]. DDS is a typical nephropathy characterized by diffuse mesangial sclerosis, genital abnormalities, and/or Wilms tumor. There is phenotypic overlap with WAGR syndrome and Frasier syndrome. Inheritance is autosomal dominant, but most cases are sporadic.
    Defects in WT1 are the cause of nephrotic syndrome type 4 (NPHS4) [MIM:256370]. A renal disease characterized clinically by proteinuria, hypoalbuminemia, hyperlipidemia and edema. Kidney biopsies show non-specific histologic changes such as focal segmental glomerulosclerosis and diffuse mesangial proliferation. Some affected individuals have an inherited steroid-resistant form and progress to end-stage renal failure. Most patients with NPHS4 show diffuse mesangial sclerosis on renal biopsy, which is a pathologic entity characterized by mesangial matrix expansion with no mesangial hypercellularity, hypertrophy of the podocytes, vacuolized podocytes, thickened basement membranes, and diminished patency of the capillary lumen.
    Defects in WT1 are a cause of Meacham syndrome (MEACHS) [MIM:608978]. Meacham syndrome is a rare sporadically occurring multiple malformation syndrome characterized by male pseudohermaphroditism with abnormal internal female genitalia comprising a uterus and double or septate vagina, complex congenital heart defect and diaphragmatic abnormalities.
    Note=A chromosomal aberration involving WT1 may be a cause of desmoplastic small round cell tumor (DSRCT). Translocation t(11;22)(p13;q12) with EWSR1.
  • Sequence similarities
    Belongs to the EGR C2H2-type zinc-finger protein family.
    Contains 4 C2H2-type zinc fingers.
  • Cellular localization
    Nucleus. Cytoplasm. Shuttles between nucleus and cytoplasm; Nucleus > nucleoplasm and Nucleus speckle.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • WIT 2 antibody
    • WT 1 antibody
    • AWT1 antibody
    • FWT1 antibody
    • GUD antibody
    • NPHS4 antibody
    • WAGR antibody
    • Wilms tumor 1 antibody
    • Wilms Tumor antibody
    • Wilms tumor protein antibody
    • Wilms' tumor gene antibody
    • Wilms' tumor protein antibody
    • WIT2 antibody
    • WT antibody
    • WT1 antibody
    • WT1_HUMAN antibody
    • WT33 antibody
    see all

Images

  • Immunohistochemistry (Frozen sections) - Anti-Wilms Tumor Protein antibody (ab15249)
    Immunohistochemistry (Frozen sections) - Anti-Wilms Tumor Protein antibody (ab15249)Image from Hartleben B et al., PLoS One. 2012;7(5):e36705. Epub 2012 May 7. Fig 3.; doi:10.1371/journal.pone.0036705; May 7, 2012, PLoS ONE 7(5): e36705.
    Immunohistochemical analysis of murine kidney tissue, staining Wilms Tumor Protein (red) with ab15249 at 1/300 dilution. A fluorescent-conjugated secondary antibody was used to detect staining.

References

This product has been referenced in:
See all 19 Publications for this product

Publishing research using ab15249? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 17 Abreviews or Q&A

IHC - Wholemount abreview for Anti-Wilms Tumor Protein antibody

 Good
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
abreview image
Application
IHC - Wholemount
Sample
Mouse Embryo (lung, mesothelial cells)
Specification
lung, mesothelial cells
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Abcam user community

Verified customer

Submitted Dec 14 2016

Immunohistochemistry (Frozen sections) abreview for Anti-Wilms Tumor Protein antibody

 Excellent
Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (lung, mesothelial cells)
Permeabilization
No
Specification
lung, mesothelial cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

Submitted Dec 14 2016

Question

This antibody did not work in Western blot as originally thought it would.

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Answer

Thank you for your phone call today, and I apologize for the troublewith this antibody.

Your credit note ID is ***

If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expertassistance or advice.

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Question

Thanks for your mail. I used two anti-WT-1 antibodies (ab15249 and ab64681) and both are giving multiple bands with my experiments. The ab64681 is giving one huge band at 100 KDa along with other additional bands that are alsotec observed with ab15249.I wrote my last mail as you asked forfeedback onyour antibodies.When I compared I find ab15249better than ab64681 with my experimental system, although it also produced multiple bands.I am still confused with the occurrence of these bands.

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Answer

Thank you for your reply. To ensure that your message gets directed to the person who has been handling your case, please do not alter the subject line when replying to our emails. This will allow your case to be tracked effectively in our system.
After reviewing the details of your contacts with my colleagues, I have only located the image of ab73466 with ab64681. It appears that when running the peptide ab73466 in SDS-PAGE, you detected a band around 75 kDa. While ab73466 is suitable for blocking the specificity of ab64681 after preincubation, it is only 18 amino acids in length and is therefore too small to be detected in SDS-PAGE.
This peptide is 70-90% pure and is supplied in a buffer that does not contain any additional proteins. Because this protein is synthetic, the 10-30% contamination will be amino acids, chemicals, and partial peptides, none of which would be expected to run at 75 kDa. The The presence of the 75 kDa band indicates that there may be some protein contamination in the loading or running buffer. This may also help to explain some of the additional bands in your western.
According to the UniProt database, there are 8 isoforms of Wilms Tumor Protein with molecular weights ranging from 34 - 57 kDa. This protein is also known to interact with other proteins and to form homodimers (which may be the band observed at 100 kDa). Using a buffer with ionic and non-ionic detergents (such as RIPA buffer) and boiling the protein in the presence of BME or DTT will help to ensure that all multimers are broken down.
I hope this helps, please let me know if you need any additional information or assistance.

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Question

I have asked for peptide sequence that was used to raise ab15249. In case it is not available, could you please provide me its sequence? I will be so thankful to you if you can provide that.

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Answer

Thank you for contacting us.

The exact immunogen sequence is confidential, but it is contained within the range between aa 389-449 of P19544 (Uniprot), isoform 1.

http://www.uniprot.org/uniprot/P19544

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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Question

I have received a product recall notification on ab15249.

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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.
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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Question

Dear Technical,
Hello Sci Supp! The below customer called in response to an email from Abcam regarding (recalls I think). She wants to order replacements. I informed her that our technical team would contact her today to get more information.
Customer Name:
Customer Email:
Abcam orders: #
Products in question:
ab15249 : Rabbit polyclonal to Wilms Tumor Protein
ab65835 : Rabbit polyclonal to ANGPT1 + ANGPT2
Thanks,

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Answer

Thank you for contacting Abcam.
For ab15429 we have found that there are problems with this antibody in western blot and that we can no longer guarantee that it will work in this application. However it is still guaranteed to work in IP and IHC-P. If you had originally purchased this antibody to work in WB and you are having problems with it, then I will be happy to replace the antibody.
As for ab65835 we have found that cross reacting with angiopoietin 1 and therefore we would be happy to either replace this antibody with another angiopoietin 1 antibody of your choice, possible ab8451 or ab94684.
I do apologize for any inconvenience these issues may have caused you and I look forward to hearing from you so that I can get this sorted out.
If there is anything I can help with, please let me know.

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Question

I checked the datasheets online, the ab96792 should work. Thank you for your help, I'll keep in mind to contact Prodotti Gianni for our next orders, Best regards,

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Answer

Thank you for your response. This is to let you know that I have confirmed and agreed on placing a new order for your customer - for one vial of ab96792 as a free of charge replacement. I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product. Please could you contact Prodotti Gianni and discuss this case with them and also get some further information about the exact shipping time. Their contact details can be found at the website: www.prodottigianni.com Merry Xmas and Happy New Year!

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Question

Thank you for your prompt reply. I haven't contacted you before because, as I said, we purchased this antibody both for immunostaining and western blotting and I've been using it so far just for stainings. Today, by change, just 1day before the 6 months from the arrival date (16/6/2011) , I checked the Scientific Support session to find more information on how to use the antibody for WB. This is the technical support answer to question number 6: " ANSWER:   Thank you for contacting Abcam. I have talked to the people in the lab and also reviewed all the data we have on ab15249 and it seems as though this antibody does not work in western blot (it gives multiple bands, which is what you are seeing). The tested in western blot application was added to the datasheet by mistake. Please accept my apologies for this mistake and I hope it has not caused you too many problems. We do have several other Wilms Tumor Protein antibodies in our catalogue and I would be very happy to send you a replacement antibody. Several antibodies that you may be interested in are: - ab64681 - ab89901 - ab96792 Once again please accept my apologies and let me know which antibody you would like to receive as a replacement. Or if you would prefer I can process a refund. I look forward to your reply. which means that the western blot application has been added to the datasheet by mistake and even if I try for months I won't have a single band on the gel and most likely this won't be related to problems with the shipping or with the storage of the product. That's why I was hoping to have it replaced with another anybody which actually works for both the applications as specified from the datasheet. Regards,  

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Answer

Thank you for getting back to me and for clarifying the situation further. It is indeed the case that after consideration Western blot application has been removed from the datasheet. Although, you have purchased this antibody via an unauthorized reseller, I could certainly offer you an alternative antibody against Wilms Tumor Protein i.e. ab89901, ab118873. Please take a look at the datasheets and also our on-line catalogue and let me know which product you would prefer to get as a free of charge replacement. It is important to mention that our authorized distributor in Italy is Prodotti Gianni SpA (www.prodottigianni.com) so we can ship the product to them - even though originally you ordered this antibody via another company. I hope this would be a satisfactory outcome for you. I look forward to receiving confirmation that you would like to proceed with a replacement vial.

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Question

Thanks for your concern. You can send me one replaced vial of ab64681 along with the blocking peptide (ab73466) urgently to execute my experimets. It would have been much better if you could send me both replaced vials since the first vial was ordered long back and I am unable to find the order copy for that now. But since I have two vials of ab15249 and therefore it is obvious that I purchased two vials from you.    Thanks,  

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Answer

Thank you for your patience in this matter. I have processed the request for you to receive ab64681 as a replacement for ab15249 which did not work in western blot as stated on the datasheet. I have requested that the peptide ab73466 be included in the order as requested so that you will be able to confirm the specificity of the bands seen in your western blot. I am continuing to look for the first order you placed for ab15249 but I wanted to get 1 vial of the new antibody and the peptide to you as soon as possible so that you can continue with your experiments. If there is anything else I can do to help, please let me know.

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1-10 of 17 Abreviews or Q&A

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