Recombinant Human Wnt-3a (High Purity) Protein

Carrier Free

Catalog # Availability Size / Price Qty
5036-WNP-010/CF

With Carrier

Catalog # Availability Size / Price Qty
5036-WNP-010
Recombinant Human Wnt-3a (High Purity) Protein Bioactivity
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Product Details
Citations (3)
FAQs
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Recombinant Human Wnt-3a (High Purity) Protein Summary

Product Specifications

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 human Wnt-3a protein
Ser19-Lys352
Accession #
N-terminal Sequence
Analysis
Ser19
Predicted Molecular Mass
37.4 kDa
SDS-PAGE
40 kDa, reducing conditions

Product Datasheets

5036-WNP (with carrier)

5036-WNP/CF (carrier free)

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

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 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.
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.

5036-WNP/CF

Formulation Lyophilized from a 0.2 μm filtered solution in PBS, NaCl, EDTA and CHAPS.
Reconstitution Reconstitute at 200 μg/mL in sterile PBS.
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.

Data Images

Bioactivity Recombinant Human Wnt-3a (High Purity) Protein Bioactivity View Larger

Recombinant Human Wnt-3a (High Purity, Catalog # 5036-WNP) induces alkaline phosphatase production by the MC3T3-E1 mouse preosteoblast cell line. The ED50 for this effect is 1.5-7.5 ng/mL.

SDS-PAGE Recombinant Human Wnt-3a (High Purity) Protein SDS-PAGE View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Citations for Recombinant Human Wnt-3a (High Purity) Protein

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.

3 Citations: Showing 1 - 3
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  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 Types: Whole Cells
    Applications: 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 Types: Recombinant Protein
    Applications: Binding Assay
  3. Commercially Available Preparations of Recombinant Wnt3a Contain Non-Wnt Related Activities Which May Activate TGF-beta Signaling.
    Authors: Carthy J, Engstrom U, Heldin C, Moustakas A
    J Cell Biochem, 0;117(4):938-45.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay

FAQs

  1. Can I reconstitute Recombinant Human Wnt-3a (High Purity) Protein (catalog # 5036-WNP) at a different concentration than recommended?

    • We cannot guarantee the performance if the protein is reconstituted at a different concentration than recommended on the product datasheet. Please contact Technical Service if you have any questions or concerns about reconstitution.

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