Recombinant Human Wnt-3a (High Purity) Protein, CF

(2 citations)   
  • Purity
    >90%, by SDS-PAGE with silver staining
  • Endotoxin Level
    <0.10 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by its ability to induce alkaline phosphatase production by MC3T3‑E1 mouse preosteoblast cells. The ED50 for this effect is 1.5-7.5 ng/mL. Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells.

    Protein concentrations should be titrated based on cell type and if appropriate, passage number of the cell line.
    Optimal concentrations should be determined by each laboratory for each application.

  • Source
    Chinese Hamster Ovary cell line, CHO-derived Ser19-Lys352
  • Accession #
  • N-terminal Sequence
    Analysis
    Ser19
  • Predicted Molecular Mass
    37.4 kDa
  • SDS-PAGE
    40 kDa, reducing conditions
Carrier Free
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
5036-WNP/CF
 
5036-WNP
Formulation Lyophilized from a 0.2 μm filtered solution in PBS, NaCl, EDTA and CHAPS.
Formulation Lyophilized from a 0.2 μm filtered solution in PBS, NaCl, EDTA and CHAPS with BSA as a carrier protein.
Reconstitution Reconstitute at 200 μg/mL in sterile PBS.
Reconstitution Reconstitute at 200 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Data Images
Recombinant Human Wnt-3a (High Purity, Catalog # 5036-WNP/CF) induces alkaline phosphatase production by the MC3T3-E1 mouse preosteoblast cell line. The ED50 for this effect is 1.5-7.5 ng/mL.
1 μg/lane of Recombinant Human Wnt-3a (High Purity) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 40 kDa.
Background: Wnt-3a

Wnt-3a is one of 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). Wnts bind to the cell surface Frizzled family receptors in conjunction with low-density lipoprotein receptor-related protein family receptors (LRP5 or 6) resulting in the stabilization of intracellular beta -catenin levels (2). As intracellular beta -catenin levels rise, beta -catenin binds to TCF/LEF transcription factors leading to expression of Wnt target genes (3). Endo-IWR 1 (Catalog # 3532, # PSM1324) is a cell-permeant small molecule inhibitor of Axin turnover that suppresses Wnt signal transduction by stabilizing the beta -catenin destruction complex (4). Wnt-3a is a 44 kDa secreted hydrophobic glycoprotein containing a conserved pattern of 24 cysteine residues (5). Wnt-3a has two N-linked glycosylation sites (Asn 87, Asn 298), and Ser 209 is modified with palmitoleic acid (6). Glycosylation and acylation are essential for efficient Wnt secretion and biological activity, respectively (6, 7). Human Wnt-3a shares 96% amino acid (aa) identity with mouse mouse, bovine and canine Wnt-3a, and 89%, 86% and 84% aa identity with chicken, Xenopus and zebrafish Wnt-3a, respectively. It also shares 87% aa identity with Wnt3. During embryonic development, Wnt-3a is necessary for proper development of the hippocampus, anterior-posterior patterning, somite development, and tailbud formation (9-12). Wnt-3a also promotes self-renewal of hematopoietic stem cells, neural stem cells, and embryonic stem cells (13, 14).

  • References:
    1. Willert, K. and Nusse, R. (2012) Cold Spring Harb. Perspect. Biol. 4:a007864.
    2. MacDonald, B.T. and X. He (2012) Cold Spring Harb. Perspect. Biol. 4:a007880.
    3. Korinek, V. et al. (1997) Science 275:1784.
    4. Chen, B. et al. (2009) Nat. Chem. Biol. 5:100.
    5. Smolich, B.D. et al. (1993) Mol. Biol. Cell 4:1267.
    6. Takada, R. et al. (2006) Dev. Cell 11:791.
    7. Komekado, H. (2007) Genes Cells 12:521.
    8. Dunty Jr. W. C. et al. (2008) Development 135:85.
    9. Ikeya, M. and S. Takada (2001) Mech. Dev. 103:27.
    10. Lee, S. M. et al. (2000) Development 127:457.
    11. Takada, S. et al. (1994) Genes Dev. 8:174.
    12. Willert, K. et al. (2003) Nature 423:6938.
    13. Kalani, M.Y. et al. (2008) Proc. Natl. Acad. Sci. USA 105:16970.
    14. Ten Berge, D. et al. (2011) Nat. Cell Biol. 13:1070.
  • 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
Citations:

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.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Wnt signaling blockage inhibits cell proliferation and migration, and induces apoptosis in triple-negative breast cancer cells.
    Authors: Bilir B, Kucuk O, Moreno C
    J Transl Med, 2013;11(0):280.
    Species: Human
    Sample Type: Whole Cells
    Application: Bioassay
  2. Reconstitution of a frizzled8.Wnt3a.LRP6 signaling complex reveals multiple Wnt and Dkk1 binding sites on LRP6.
    Authors: Bourhis E, Tam C, Franke Y, Bazan JF, Ernst J, Hwang J, Costa M, Cochran AG, Hannoush RN
    J. Biol. Chem., 2010;285(12):9172-9.
    Species: Human
    Sample Type: Recombinant Protein
    Application: Binding Assay

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