Human/Mouse Wnt-3a Antibody

Recombinant Monoclonal Antibody..
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects mouse Wnt-3a in direct ELISAs.
  • Source
    Recombinant Monoclonal Rat IgG2A Clone # 217804.2R
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    E.coli-derived recombinant mouse Wnt-3a
    Ser36-Gln75, Trp219-Arg269
    Accession # P27487
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Mouse Wnt‑3a by Western Blot. Western blot shows lysates of CHO Chinese hamster ovary cell line either mock transfected or transfected with mouse Wnt-3a. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human/Mouse Wnt‑3a Monoclonal Antibody (Catalog # MAB13242) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Wnt‑3a at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Intracellular Staining by Flow Cytometry
Detection of Wnt3A in BG01V Human Stem Cells by Flow Cytometry. BGO1v human embryonic stem cells were stained with Rat Anti-Human/Mouse Wnt-3a Monoclonal Antibody (Catalog # MAB1324, filled histogram) or isotype controlantibody (Catalog # MAB006, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed and permeabilized with  FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Wnt-3a

Wnt-3a is one of about 19 vertebrate members of the Wingless-type MMTV integration site (Wnt) family of highly conserved cysteine-rich secreted glycoproteins important for normal developmental processes (1‑3). Wnts bind to receptors of the Frizzled family in conjunction with a coreceptor of the low-density lipoprotein receptor-related protein family (LRP-5 or -6), or the Ryk atypical receptor tyrosine kinase (1, 4). Mouse Wnt-3a is a 44 kDa secreted hydrophobic glycoprotein containing a conserved pattern of 24 cysteine residues (5). Like other Wnts, Wnt-3a is modified by palmitate addition (at Cys 77) following glycosylation, which increases its hydrophobicity, secretion and activity (6, 7). A second site at Ser 209 is modified by palmitoleic acid and also contributes to activity and secretion (8). Mouse Wnt-3a shares 96% amino acid (aa) identity with human Wnt-3a, and 97% with bovine and canine Wnt-3a. The rat Wnt-3a precursor as it is apparently translated shares 100% aa identity with mouse Wnt-3a aa 63-352 (9). Wnt-3a also shares 87% aa identity with Wnt-3. During development, Wnt-3a is morphogen that is thought to coordinate somitogenesis and mesoderm boundary determination, and is expressed at the same locations and times as Wnt-2b and Wnt-5a (10). When Wnt-3a is deleted, mice fail to develop a hippocampus, and show defects in anterior-posterior patterning, somite development and tailbud formation (10-13). Recombinant Wnt-3a promotes proliferation of committed stem cell lineages in vitro, and may help maintain the cells in an undifferentiated state (6, 14) For example, Wnt-3a can induce self-renewal of hematopoietic stem cells, allowing expansion without further differentiation (6).

  • References:
    1. Mikels, A.J. and R. Nusse (2006) Oncogene 25:7461.
    2. Miller, J.R. (2001) Genome Biol. 3:3001.
    3. Roelink, H. and R. Nusse (1991) Genes Dev. 5:381.
    4. Lu, W et al. (2004) Cell 119:97.
    5. Smolich, B.D. et al. (1993) Mol. Biol Cell 4:1267.
    6. Willert, K. et al. (2003) Nature 423:448.
    7. Komekado, H. et al. (2007) Genes Cells 12:521.
    8. Takada, R. et al. (2006) Dev. Cell 11:791.
    9. Entrez Accession # NP_001100475.
    10. Dunty, W.C. et al. (2008) Development 135:85.
    11. Lee, S.M.K. et al. (2000) Development 127:457.
    12. Takada, S. et al. (1994) Genes Dev. 8:174.
    13. Ikeya, M. and S. Takada (2001) Mech. Dev. 103:27.
    14. Nusse, R. (2008) Cell Res. 18:523.
  • Long Name:
    Wingless-type MMTV Integration Site Family, Member 3a
  • Entrez Gene IDs:
    89780 (Human); 22416 (Mouse)
  • Alternate Names:
    MGC119418; MGC119419; MGC119420; protein Wnt-3a; wingless-type MMTV integration site family, member 3A; Wnt3a; Wnt-3a
Related Research Areas
Isotype Controls
Description Application Cat# Citations Images  

Rat IgG2A Isotype Control

Ctrl MAB006 58  
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Normal Rat IgG Control

Ctrl 6-001-A 7
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Normal Rat IgG Control (Azide Free)

Ctrl 6-001-F 1
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Molecular Weight Markers
Description Application Cat# Citations Images  

Full-Range Molecular Weight Marker, Biotinylated

MW001
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Secondary Antibodies
Description Application Cat# Citations Images  

Rat IgG Horseradish Peroxidase-conjugated Antibody

WB, Simple Western HAF005 3  
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Goat Anti-Rat F(ab)2 IgG (H+L)
APC-conjugated Antibody

Flow F0113 1  
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Rat F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0105B 2  
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Goat Anti-Rat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL013 2
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Goat Anti-Rat IgG Antibody

WB AF005 1
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Goat Anti-Rat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL014
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Goat Anti-Rat IgG Biotinylated Antibody

WB BAF005 1
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Rat F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0104B  
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Goat Anti-Rat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL015
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Goat Anti-Rat F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0115
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