Human/Mouse Wnt-3a Antibody

MAB1324 has been discontinued and is replaced by MAB9025.
(7 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects mouse Wnt-3a in direct ELISAs and human and mouse Wnt-3a in Western blots. In direct ELISAs and Western blots, this antibody does not cross-react with recombinant mouse Wnt-1, -4, or 5a.
  • Source
    Monoclonal Rat IgG2A Clone # 217804
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant mouse Wnt‑3a
    Ser36-Gln75 and Trp219-Arg269
    Accession # P27467.1
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
  • Western Blot
    2 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
  • Neutralization
    Measured by its ability to neutralize Wnt‑3a-induced alkaline phosphatase production in the MC3T3‑E1 mouse preosteoblast cell line. The Neutralization Dose (ND50) is typically 3-15 µg/mL in the presence of 10 ng/mL Recombinant Mouse Wnt‑3a or 20 ng/mL Recombinant Human Wnt‑3a.
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 Human and Mouse Wnt‑3a by Western Blot. Western blot shows lysates of BG01V human embryonic stem cells and mouse lung tissue. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human/Mouse Wnt‑3a Monoclonal Antibody (Catalog # MAB1324) 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 Wnt-3a in BG01V Human Stem Cells by Flow Cytometry. BG01V human embryonic stem cells were stained with Rat Anti-Human/Mouse Wnt-3a Monoclonal Antibody (Catalog # MAB1324, filled histogram) or isotype control antibody (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).
Alkaline Phosphatase Production Induced by
Wnt‑3a and Neutralization by Human/Mouse Wnt‑3a Antibody.
Recombinant Mouse Wnt‑3a (Catalog # 1324-WN) induces alkaline phosphatase production in the MC3T3‑E1 mouse preosteoblast cell line in a dose-dependent manner (orange line). Under these conditions, alkaline phosphatase production elicited by Recombinant Mouse Wnt‑3a (10 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Human/Mouse Wnt‑3a Monoclonal Antibody (Catalog # MAB1324). The ND50 is typically 3-15 µg/mL.
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
Related Research Areas

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

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Sample Type
  1. SULF2 overexpression positively regulates tumorigenicity of human prostate cancer cells.
    Authors: Vicente C, Lima M, Nader H, Toma L
    J Exp Clin Cancer Res, 2015;34(0):25.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  2. (Pro)renin receptor is crucial for Wnt/beta-catenin-dependent genesis of pancreatic ductal adenocarcinoma.
    Authors: Shibayama Y, Fujimori T, Nguyen G, Hirose T, Totsune K, Ichihara A, Kitada K, Nakano D, Kobori H, Kohno M, Masaki T, Suzuki Y, Yachida S, Nishiyama A
    Sci Rep, 2015;5(0):8854.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Drugging a stem cell compartment using Wnt3a protein as a therapeutic.
    Authors: Dhamdhere, Girija R, Fang, Mark Y, Jiang, Jie, Lee, Katherin, Cheng, Du, Olveda, Rebecca, Liu, Bo, Mulligan, Kimberle, Carlson, Jeffery, Ransom, Ryan C, Weis, William, Helms, Jill A
    PLoS ONE, 2014;9(1):e83650.
    Species: Human
    Sample Type: recombinant protein
    Application: WB
  4. Impaired skeletal muscle regeneration in the absence of fibrosis during hibernation in 13-lined ground squirrels.
    Authors: Andres-Mateos, Eva, Mejias, Rebeca, Soleimani, Arshia, Lin, Brian M, Burks, Tyesha N, Marx, Ruth, Lin, Benjamin, Zellars, Richard, Zhang, Yonggang, Huso, David L, Marr, Tom G, Leinwand, Leslie A, Merriman, Dana K, Cohn, Ronald D
    PLoS ONE, 2012;7(11):e48884.
    Species: Squirrels
    Sample Type: Tissue Homogenates
    Application: WB
  5. Cooperative folding and ligand-binding properties of LRP6 beta-propeller domains.
    Authors: Liu CC, Pearson C, Bu G
    J. Biol. Chem., 2009;284(22):15299-307.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  6. Prostate tumor progression is mediated by a paracrine TGF-beta/Wnt3a signaling axis.
    Authors: Li X, Placencio V, Iturregui JM, Uwamariya C, Sharif-Afshar AR, Koyama T, Hayward SW, Bhowmick NA
    Oncogene, 2008;27(56):7118-30.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  7. Nephroblastoma overexpressed (Nov) inhibits osteoblastogenesis and causes osteopenia.
    Authors: Rydziel S, Stadmeyer L, Zanotti S, Durant D, Smerdel-Ramoya A, Canalis E
    J. Biol. Chem., 2007;282(27):19762-72.
    Species: Mouse
    Sample Type: GST and GST-Nov Fusion proteins
    Application: WB

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