Recombinant Human Wnt-3a (High Purity) Protein

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.

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.

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human Wnt-3a (High Purity) Protein Summary

• Details of Functionality: 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 #: NP_149122
• N-terminal Sequence: Ser19
• Protein/Peptide Type: Recombinant Proteins
• Gene: WNT3A
• Purity: >90%, by SDS-PAGE with silver staining
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 37.4 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 40 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS, NaCl, EDTA and CHAPS with BSA as a carrier protein.
• Purity: >90%, by SDS-PAGE with silver staining
• Reconstitution Instructions: Reconstitute at 200 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

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

• MGC119418
• MGC119419
• MGC119420
• protein Wnt-3a
• wingless-type MMTV integration site family, member 3A
• Wnt3a
• Wnt-3a

Background

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).

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.

Publications for Wnt-3a (5036-WNP)(4)

Publications using 5036-WNP

• J Krumm, K Sekine, P Samaras, A Brazovskaj, M Breunig, R Yasui, A Kleger, H Taniguchi, M Wilhelm, B Treutlein, JG Camp, B Kuster High temporal resolution proteome and phosphoproteome profiling of stem cell-derived hepatocyte development Cell Reports, 2022-03-29;38(13):110604. 2022-03-29 [PMID: 35354033] (Bioassay, Human)
• Carthy J, Engstrom U, Heldin C, Moustakas A Commercially Available Preparations of Recombinant Wnt3a Contain Non-Wnt Related Activities Which May Activate TGF-beta Signaling. J Cell Biochem, 2015-09-22;117(4):938-45. 2015-09-22 [PMID: 26369756] (Bioassay, Mouse)
• Bilir B, Kucuk O, Moreno C Wnt signaling blockage inhibits cell proliferation and migration, and induces apoptosis in triple-negative breast cancer cells. J Transl Med, 2013-11-04;11(0):280. 2013-11-04 [PMID: 24188694] (Bioassay, Human)
• Bourhis E, Tam C, Franke Y, Bazan JF, Ernst J, Hwang J, Costa M, Cochran AG, Hannoush RN Reconstitution of a frizzled8.Wnt3a.LRP6 signaling complex reveals multiple Wnt and Dkk1 binding sites on LRP6. J. Biol. Chem., 2010-01-21;285(12):9172-9. 2010-01-21 [PMID: 20093360] (Binding Assay, Human)

Bioinformatics

• Gene Symbol: WNT3A
• Uniprot:
  • NP_149122(Human)