RecombinantRabMAb

Recombinant Anti-Wnt3a antibody [EPR21889] (ab219412)

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Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Flow Cytometry (Intracellular) - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Immunoprecipitation - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Immunocytochemistry/ Immunofluorescence - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
  • Anti-Wnt3a antibody [EPR21889] (ab219412)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21889] to Wnt3a
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

Overview

  • Product name

    Anti-Wnt3a antibody [EPR21889]
    See all Wnt3a primary antibodies

  • Description

    Rabbit monoclonal [EPR21889] to Wnt3a

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: L Wnt-3a (Wnt-3a stably transfected Mouse subcutaneous connective tissue areolar and adipose fibroblast) supernatant and whole cell lysate. His-tagged human Wnt3a recombinant protein (aa19-352). Human fetal brain and fetal skin lysate. HCT 116 whole cell lysate. IHC-P: Mouse and rat E14.5 tissue. ICC/IF: L Wnt-3a cells. Flow Cyt (intra): L Wnt-3a cells. IP: L Wnt-3a whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219412 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
Flow Cyt (Intra)
1/500.
WB
1/1000. Predicted molecular weight: 39 kDa.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP
1/30.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Predicted molecular weight: 39 kDa.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP
1/30.

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.

  • Tissue specificity

    Moderately expressed in placenta and at low levels in adult lung, spleen, and prostate.

  • Sequence similarities

    Belongs to the Wnt family.

  • Cellular localization

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

  • Database links

    • Alternative names

      • Protein Wnt 3a Precursor antibody
      • Protein Wnt-3a antibody
      • Wingless type MMTV integration site family member 3A antibody
      • Wnt 3a antibody
      • wnt3a antibody
      • WNT3A protein antibody
      • WNT3A_HUMAN antibody
      see all

    Images

    • Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
      All lanes : Anti-Wnt3a antibody [EPR21889] (ab219412) at 1/1000 dilution

      Lane 1 : Human fetal brain lysate
      Lane 2 : Human fetal skin lysate
      Lane 3 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 39 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

    • Flow Cytometry (Intracellular) - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Flow Cytometry (Intracellular) - Anti-Wnt3a antibody [EPR21889] (ab219412)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) cell line (right) labeling Wnt3a with ab219412 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) (left). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

      Two Wnt3a-expressing populations can be seen labeled in this stably transfected cell line.

    • Immunoprecipitation - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Immunoprecipitation - Anti-Wnt3a antibody [EPR21889] (ab219412)

      Wnt3a was immunoprecipitated from 0.35 mg L Wnt-3a (Wnt-3a stably-transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) whole cell lysate with ab219412 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219412 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
      Lane 1: L Wnt-3a (Wnt-3a stably-transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) whole cell lysate 10 µg (Input).
      Lane 2: ab219412 IP in L Wnt-3A whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219412 in L Wnt-3a whole cell lysate.
      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 10 seconds.

    • Immunocytochemistry/ Immunofluorescence - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Immunocytochemistry/ Immunofluorescence - Anti-Wnt3a antibody [EPR21889] (ab219412)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) cells labeling Wnt3a with ab219412 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in L Wnt-3A cell line. The nuclear counter stain is DAPI (blue).
      Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
      The negative control is the secondary antibody only.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)

      Immunohistochemical analysis of paraffin-embedded rat E14.5 tissue labeling Wnt3a with ab219412 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on the hem remnant of rat E14.5 (PMID: 15509764; PMID: 16410414) is observed. Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)

      Immunohistochemical analysis of paraffin-embedded mouse E14.5 tissue labeling Wnt3a with ab219412 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on the hem remnant of mouse E14.5 (PMID: 15509764; PMID: 16410414) is observed. Counter stained with Hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
      All lanes : Anti-Wnt3a antibody [EPR21889] (ab219412) at 1/20000 dilution

      Lane 1 : His-tagged human Wnt3 recombinant protein (aa22-355), 40 ng
      Lane 2 : His-tagged human Wnt3a recombinant protein (aa19-352), 20 ng

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 39 kDa


      Exposure time: 3 minutes


      Blocking/ Dilution buffer and concentration: 5% NFDM/TBST.

    • Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
      Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)
      All lanes : Anti-Wnt3a antibody [EPR21889] (ab219412) at 1/1000 dilution

      Lane 1 : L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) supernatant at 20 µl
      Lane 2 : L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) whole cell lysate at 20 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 39 kDa



      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

      Exposure times: Lane 1: 3 minutes; Lane 2: 3 seconds.

    • Anti-Wnt3a antibody [EPR21889] (ab219412)
      Anti-Wnt3a antibody [EPR21889] (ab219412)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (15)

    Publishing research using ab219412? Please let us know so that we can cite the reference in this datasheet.

    ab219412 has been referenced in 15 publications.

    • Xu H  et al. IMPDH2 promotes cell proliferation and epithelial-mesenchymal transition of non-small cell lung cancer by activating the Wnt/ß-catenin signaling pathway. Oncol Lett 20:219 (2020). PubMed: 32963625
    • Sun H  et al. Hsa_circ_0001649 restrains gastric carcinoma growth and metastasis by downregulation of miR-20a. J Clin Lab Anal 34:e23235 (2020). PubMed: 32212290
    • Chen X  et al. Long non-coding RNA GHET1 promotes osteosarcoma development and progression via Wnt/ß-catenin signaling pathway. Oncol Rep 44:349-359 (2020). PubMed: 32319657
    • Gao Y  et al. Wnt3a Inhibitor Attenuates Remifentanil?-Induced Hyperalgesia via Downregulating Spinal NMDA Receptor in Rats. J Pain Res 13:1049-1058 (2020). PubMed: 32547170
    • Li K & Yuan C MicroRNA-103 modulates tumor progression by targeting KLF7 in non-small cell lung cancer. Int J Mol Med 46:1013-1028 (2020). PubMed: 32582959
    View all Publications for this product

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