Overview

  • Product name
  • Description
    Rabbit polyclonal to Wnt3a
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Fr, IP, IHC - Wholemount, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Chicken, Human, Zebrafish
    Predicted to work with: Xenopus laevis, a wide range of other species
  • Immunogen

    Synthetic peptide derived from within residues 250 - 350 of Human Wnt3a.

    Read Abcam's proprietary immunogen policy (Peptide available as ab23327.)

  • Positive control
    • In WB, this antibody gave a positive signal when tested against recombinant Wnt3a protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab28472 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. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).Can be blocked with Human Wnt3a peptide (ab23326) or Human Wnt3a peptide (ab23327).
IHC-Fr Use at an assay dependent concentration. PubMed: 21048974
IP Use at an assay dependent concentration. PubMed: 21740904
IHC - Wholemount Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    Images

    • Anti-Wnt3a antibody (ab28472) at 1 µg/ml + Mouse Wnt3a Full-length Recombinant Protein (Tagged) at 0.1 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Predicted band size: 39 kDa
      Observed band size: 68 kDa
      why is the actual band size different from the predicted?



      ab28472 was tested against Mouse Wnt3a Full-length Recombinant Protein predicted to run at the molecular weight of 68kDa.

    • ab28472 stained human embryonic stem cells differentiated into a heterogeneous mix of cells at 5 ug/ml.
    • ab28472 staining Wnt3a in the dentate gyrus of rat bain tissue by Immunohistochemistry (Frozen sections).
      Brains were sectioned in the coronal plane at 20 µm. The slides were left in a fume hood to dry overnight, then stored at -70°C. For each rat twelve sections through the dorsal hippocampus were analyzed. For single staining with Wnt3a, sections were incubated with ab28472 at 5 ug/ml in 0.5% Triton with 1% goat serum) overnight in a humidified chamber at room temperature. The following day, after two washes with KPBS, they were incubated in Alexa Fluor 568 goat anti-rabbit (1/200 in 0.5% Triton) for one hour, and washed twice with KPBS and covered slipped.
    • IHC-Wholemount image of Wnt3a (ab28472) on zebrafish larvae. Whole zebrafish larvae were fixed in paraformaldehyde and then permeabilized in proteinase K, before blocking in 5% serum for 1 hour at RT. The image above is a ventral view of the head, showing the ceratohyal cartilage (of the jaw).

      See Abreview

    References

    This product has been referenced in:
    • Zhao SJ  et al. SPARCL1 suppresses osteosarcoma metastasis and recruits macrophages by activation of canonical WNT/ß-catenin signaling through stabilization of the WNT-receptor complex. Oncogene 37:1049-1061 (2018). Read more (PubMed: 29084211) »
    • Hu J  et al. Morin inhibits proliferation and self-renewal of CD133+ melanoma cells by upregulating miR-216a. J Pharmacol Sci 136:114-120 (2018). WB . Read more (PubMed: 29496393) »
    See all 30 Publications for this product

    Customer reviews and Q&As

    1-10 of 11 Abreviews or Q&A

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (skin)
    Permeabilization
    Yes - triton
    Specification
    skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Formaldehyde

    Dr. Tina Yuan

    Verified customer

    Submitted Sep 10 2018

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (E8.5 mouse embryo sections)
    Specification
    E8.5 mouse embryo sections
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - PBST (PBS / 0.5% v/v Triton X-100 (Sigma))
    Blocking step
    NOVOCASTRA protein block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Nov 16 2012

    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this Wnt3a antibody.

    The details you have kindly provided will enable us to investigate this case for you and also gives us vital information for our monitoring of product quality.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Although post-translational modifications such as palmitoylation or glycosylation (as described for Wnt3a) can alter the electrophoretic mobility of the protein so that it runs at a higher molecular weight, I am not sure whether this is the cause of the problem you observed. Also, the protocol seems absolutely fine to me andI am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial.

    I apologise for the inconvenience and am pleased to offer you a free of charge replacement. The replacement could be a new vial of ab28472 or our alternative Wnt3a antibody ab19925 or any other primary antibody of your choice. Otherwise, I would be pleased to arrange acredit note in compensation.

    In order to arrange this, could you please let me know one of the following: the order number, the PO number, the date of purchase or at least the address of your lab?

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More
    Abreviews
    Application
    IHC - Wholemount
    Sample
    Zebrafish Embryo (whole embryo - 24hpf)
    Specification
    whole embryo - 24hpf

    Ms. Emerald Butko

    Verified customer

    Submitted Mar 09 2012

    Answer

    Thank you for contacting Abcam.

    The discount code has been added to your Abreview.

    If there is anything else I can help you with, please let me know.

    Read More
    Application
    Western blot
    Sample
    Guinea pig Cell lysate - whole cell (brain)
    Loading amount
    30 µg
    Specification
    brain
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Feb 29 2012

    Answer

    Thank you for your enquiry and interest.

    I regret to inform you that we are unable to change the formulation of this antibody.

    Apologies! If you need any further assistance in the future, please do not hesitate to contact me.

    Read More

    Answer

    Thank you for contacting us and I apologize that you did not receive these codes earlier. These codes will each give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for each antibody in guinea pig 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. Each 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 Abreviews and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.  

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    IHC - Wholemount
    Sample
    Zebrafish Embryo (Whole mount (whole zebrafish larvae))
    Specification
    Whole mount (whole zebrafish larvae)

    Dr. Chrissy Hammond

    Verified customer

    Submitted Oct 24 2011

    Answer

    Thank you for your enquiry. I would like to align the immunogen peptide with the canine Wnt3a sequence but I have been unable to locate the canine Wnt3a sequence. If you have the canine Wnt3a sequence, here is the immunogen sequence used to produce ab28472 which you could use to determine theoretical likelihood of cross-reactivity. The data regarding utility of ab28472 for IHC-P is confusing. The Abreview data notes an 'inconclusive' rating from the researcher. And there is a Reference for this antibody that appears to have successfully used ab28472 in IHC-P: Gerhart J et al. Myo/Nog cell regulation of bone morphogenetic protein signaling in the blastocyst is essential for normal morphogenesis and striated muscle lineage specification. Dev Biol : (2011). IHC-P; Chicken. PubMed: 21884693 So I would say this antibody may still be useful for IHC-P.

    Read More

    1-10 of 11 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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