Overview

  • Product name

  • Description

    Rabbit polyclonal to Wnt3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rabbit, Chicken
  • Immunogen

    Synthetic peptide corresponding to Mouse Wnt3 aa 250-350 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab32248)

  • Positive control

    • A549 (Human Lung Carcinoma) Whole Cell Lysate Human lung adenocarcinoma FFPE tissue sections

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32249 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 37 kDa (predicted molecular weight: 40 kDa).
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Target

  • Function

    Ligand for members of the frizzled family of seven transmembrane receptors. Wnt-3 and Wnt-3a play distinct roles in cell-cell signaling during morphogenesis of the developing neural tube.
  • Involvement in disease

    Defects in WNT3 are the cause of autosomal recessive tetra-amelia (ARTTRA) [MIM:273395]. Tetra-amelia is a rare human genetic disorder characterized by complete absence of all four limbs and other anomalies such as craniofacial, nervous system, pulmonary, skeletal and urogenital defects.
  • Sequence similarities

    Belongs to the Wnt family.
  • Cellular localization

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

  • Alternative names

    • Int 4 antibody
    • INT4 antibody
    • MGC131950 antibody
    • MGC138321 antibody
    • MGC138323 antibody
    • Proto-oncogene Int-4 homolog antibody
    • Proto-oncogene Wnt-3 antibody
    • wingless type MMTV integration site family member 3 antibody
    • Wnt 3 proto oncogene protein antibody
    • WNT 3 proto oncogene protein precursor antibody
    • wnt3 antibody
    • WNT3_HUMAN antibody
    see all

Images

  • All lanes : Anti-Wnt3 antibody (ab32249) at 1 µg/ml

    Lane 1 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 2 : A549 (Human lung carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 3 : Wnt3a Recombinant Protein at 0.01 µg

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

    Performed under reducing conditions.

    Predicted band size: 40 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 50 kDa (possible cross reactivity)



    ab32249 is not expected to recognize Wnt3a due to the immunogen having low sequence homology.
  • Wnt3 was immunoprecipitated using 0.5mg MCF 7 whole cell extract, 5µg of Rabbit polyclonal to Wnt3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, MCF 7 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32249.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 40kDa: Wnt3.
  • IHC image of ab32249 staining in human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab32249, 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.

References

This product has been referenced in:

  • Wang J  et al. (Pro)renin receptor promotes colorectal cancer through the Wnt/beta-catenin signalling pathway despite constitutive pathway component mutations. Br J Cancer 120:229-237 (2019). Read more (PubMed: 30555158) »
  • Pouyafar A  et al. Treatment of cancer stem cells from human colon adenocarcinoma cell line HT-29 with resveratrol and sulindac induced mesenchymal-endothelial transition rate. Cell Tissue Res N/A:N/A (2019). Read more (PubMed: 30758710) »
See all 13 Publications for this product

Customer reviews and Q&As

Answer

Thank you for contacting us. Here are the links to our Whole mount staining protocols - Protocols and troubleshooting tips for whole mount immunohistochemical staining: https://www.abcam.com/index.html?pageconfig=resource&rid=11330 and for the mouse staining in particular (also attached): https://www.abcam.com/ps/pdf/protocols/whole_mount_mouse_chick.pdf Please let me know if you like to receive the testing discounts for the 3 antibodies. Here is some more information in that regard: For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. 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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