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    wnt3a-antibody-epr21889-ab219412.pdf

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RecombinantRabMAb

Recombinant Anti-Wnt3a antibody [EPR21889] (ab219412)

  • Datasheet
  • SDS
  • Certificate of Compliance
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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 APC Carrier Free HRP PE

You may also be interested in

Biochemical
Product image
C59, Wnt Antagonist, Wnt signaling pathway inhibitor (ab142216)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Primary
Product image
Anti-Wnt3 and Wnt3a antibody [EPR4920(2)] (ab172612)

View more associated products

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

  • Form

    Liquid
  • Storage instructions

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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21889
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Stem Cells
    • Signaling Pathways
    • Wnt
    • Secreted
    • Developmental Biology
    • Embryogenesis
    • Morphogens

Associated products

  • Alternative Versions

    • Anti-Wnt3a antibody [EPR21889] - BSA and Azide free (ab234099)
    • Alexa Fluor® 647 Anti-Wnt3a antibody [EPR21889] (ab270093)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunohistochemistry reagents

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant mouse Wnt3a protein (ab81484)
  • Related Products

    • Anti-Wnt3 and Wnt3a antibody [EPR4920(2)] (ab172612)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

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.
  • Target information above from: UniProt accession P56704 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 89780 Human
    • Entrez Gene: 22416 Mouse
    • Entrez Gene: 303181 Rat
    • Omim: 606359 Human
    • SwissProt: P56704 Human
    • SwissProt: P27467 Mouse
    • Unigene: 336930 Human
    • Unigene: 1367 Mouse
    • 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)

      Intracellular 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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (20)

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

    ab219412 has been referenced in 20 publications.

    • Yang X  et al. WNT/ß-catenin-suppressed FTO expression increases m6A of c-Myc mRNA to promote tumor cell glycolysis and tumorigenesis. Cell Death Dis 12:462 (2021). PubMed: 33966037
    • Shen X  et al. lncRNA NEAT1 facilitates the progression of colorectal cancer via the KDM5A/Cul4A and Wnt signaling pathway. Int J Oncol 59:N/A (2021). PubMed: 34109988
    • Liu C  et al. Tumor-associated macrophage-derived transforming growth factor-ß promotes colorectal cancer progression through HIF1-TRIB3 signaling. Cancer Sci 112:4198-4207 (2021). PubMed: 34375482
    • Lu X  et al. Modulation of HOXA9 after skeletal muscle denervation and reinnervation. Am J Physiol Cell Physiol 318:C1154-C1165 (2020). PubMed: 32233950
    • Chen H  et al. GPX8 is transcriptionally regulated by FOXC1 and promotes the growth of gastric cancer cells through activating the Wnt signaling pathway. Cancer Cell Int 20:596 (2020). PubMed: 33317536
    View all Publications for this product

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