Recombinant
RabMAb

Recombinant Anti-Wnt2/IRP antibody [EPR3101(2)] (ab109222)

Overview

  • Product name

    Anti-Wnt2/IRP antibody [EPR3101(2)]
    See all Wnt2/IRP primary antibodies
  • Description

    Rabbit monoclonal [EPR3101(2)] to Wnt2/IRP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Wnt2/IRP aa 250-350. The exact sequence is proprietary.
    (Peptide available as ab197186)

  • Positive control

    • Jurkat,MCF-7,T47-D, and SKBR-3 cell lysates Human fetal lung tissue
  • General notes

     

     

    Previously labelled as Wnt2. 

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109222 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 1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 40 kDa).

Target

  • Function

    Ligand for members of the frizzled family of seven transmembrane receptors. Probable developmental protein. May be a signaling molecule which affects the development of discrete regions of tissues. Is likely to signal over only few cell diameters.
  • Tissue specificity

    Expressed in brain in the thalamus, in fetal and adult lung and in placenta.
  • Sequence similarities

    Belongs to the Wnt family.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Database links

  • Form

    The WNT gene family consists of structurally related genes which encode secreted signaling proteins. These proteins have been implicated in oncogenesis and in several developmental processes, including regulation of cell fate and patterning during embryogenesis.
  • Alternative names

    • Int 1 related protein antibody
    • Int-1-like protein 1 antibody
    • Int-1-related protein antibody
    • INT1L 1 antibody
    • INT1L1 antibody
    • IRP antibody
    • IRP protein antibody
    • ONCOGENE INT1 LIKE 1 antibody
    • Protein Wnt-2 antibody
    • Secreted growth factor antibody
    • Wingless type MMTV integration site family member 2 antibody
    • WNT 2 antibody
    • WNT2 antibody
    • WNT2_HUMAN antibody
    see all

Images

  • Anti-Wnt2/IRP antibody [EPR3101(2)] (ab109222) at 1/5000 dilution (purified) + Mouse brain tissue lysate at 10 µg

    Secondary
    HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 40 kDa
    Observed band size: 34 kDa
    why is the actual band size different from the predicted?



    Blocking Buffer: 5% NFDM/TBST

    Dilution Buffer: 5% NFDM/TBST

  • Anti-Wnt2/IRP antibody [EPR3101(2)] (ab109222) at 1/1000 dilution (purified) + Rat brain tissue lysate at 20 µg

    Secondary
    HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 40 kDa
    Observed band size: 34 kDa why is the actual band size different from the predicted?



    Blocking Buffer: 5% NFDM/TBST

    Dilution Buffer: 5% NFDM/TBST

  • All lanes : Anti-Wnt2/IRP antibody [EPR3101(2)] (ab109222) at 1/5000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : SKBR-3 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 40 kDa
    Observed band size: 34 kDa why is the actual band size different from the predicted?



    Blocking Buffer: 5% NFDM/TBST

    Dilution Buffer: 5% NFDM/TBST

  • All lanes : Anti-Wnt2/IRP antibody [EPR3101(2)] (ab109222) at 1/1000 dilution (unpurified)

    Lane 1 : Jurkat cell lysates
    Lane 2 : MCF-7 cell lysates
    Lane 3 : T47-D cell lysates
    Lane 4 : SKBR-3 cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 40 kDa
    Observed band size: 34 kDa why is the actual band size different from the predicted?

References

This product has been referenced in:

  • Bläuer M  et al. Wnt/ß-catenin signalling plays diverse functions during the process of fibrotic remodelling in the exocrine pancreas. Pancreatology 19:252-257 (2019). Read more (PubMed: 30792046) »
  • Xie B  et al. Benzyl isothiocyanate suppresses development and metastasis of murine mammary carcinoma by regulating the Wnt/ß-catenin pathway. Mol Med Rep 20:1808-1818 (2019). Read more (PubMed: 31257529) »
See all 7 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you very much for providing the images and protocol info.

I have checked different publications and databases; all show cytoplasmic staining. The links to the few are attached below;

http://www.proteinatlas.org/ENSG00000105989

http://cebp.aacrjournals.org/content/20/5/1021.abstract

http://www.nature.com/cgt/journal/v16/n4/full/cgt200878a.html

The following Abcam antibody also show cytoplasmic staining

https://www.abcam.com/WNT2-antibody-ab27794.html#WNT2-Primary-antibodies-ab27794-3.jpg

I would suggest tweaking a protocol and then checking if the result improve

- Fix cell in acetone for 10 minutes, or 4% icecold PFA or icecold methanol
- Use 5 - 10% NHS for blocking (1hour)
- Try the normal ICC/IF procedure using fluorescent secondary antibody
- Use PBST (0.1% Tween) instead of PBS
-Try no primary control also

The other question I have is
- Have you stained nucleus by any DNA dye?

I hope the results will improve after following these suggestions. Please let me know if you have any feedback or would like to discuss this further.

Read More

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. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

PS: Please either fill in the attached questionnaire or attach the protocol you have been using.

Read More

Answer

Thank you for contacting Abcam. The peptide maps between amino acids ********* of human Wnt2 (UniProt:  P09544). We have not yet tested whether this antibody will react with other WNT family members, but I did Blast the immunogen sequence and it did not pull out any other Wnt family members. If there is anything else I can help you with, please let me kno

Read More

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