Recombinant Mouse Wnt-3a, Biotinylated Protein, CF

Recombinant Biotinylated Mouse Wnt-3a (Catalog # BT1324/CF) binds to Recombinant Human Frizzled‑5 Fc Chimera (Catalog # 1617-FZC) with an ED50 of 2-12 ng/mL.

• Reactivity: Mu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Mouse Wnt-3a, Biotinylated Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. In a 100 µL reaction mixture containing biotinylated Recombinant Mouse Wnt-3a at 50 ng/mL and Recombinant Human Frizzled‑5 Fc Chimera (Catalog # 1617-FZC) dilutions, the concentration of Recombinant Human Frizzled‑5 Fc Chimera that produces 50% of the maximal binding response is 2-12 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 mouse Wnt-3a protein
  Ser19-Lys352
• Accession #: P27467
• N-terminal Sequence: Ser19
• Structure / Form: Biotinylated protein via amines
• Protein/Peptide Type: Recombinant Proteins
• Purity: >75%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 37 kDa (unlabeled).
  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.

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, EDTA and CHAPS.
• Purity: >75%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Mouse Wnt-3a, Biotinylated Protein, CF

• 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). 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 shares 100% aa identity with mouse Wnt-3a aa 63-352 (8), and 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 (13-15).

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. Entrez Accession # NP_001100475.
9. Dunty Jr. W. C. et al. (2008) Development 135:85.
10. Ikeya, M. and S. Takada (2001) Mech. Dev. 103:27.
11. Lee, S. M. et al. (2000) Development 127:457.
12. Takada, S. et al. (1994) Genes Dev. 8:174.
13. Willert, K. et al. (2003) Nature 423:6938.
14. Kalani, M.Y. et al. (2008) Proc. Natl. Acad. Sci. USA 105:16970.
15. Ten Berge, D. et al. (2011) Nat. Cell Biol. 13:1070.

Publications for Wnt-3a (BT1324/CF)(1)

Publication using BT1324/CF

• J Yun, S Hansen, O Morris, DT Madden, CP Libeu, AJ Kumar, C Wehrfritz, AH Nile, Y Zhang, L Zhou, Y Liang, Z Modrusan, MB Chen, CC Overall, D Garfield, J Campisi, B Schilling, RN Hannoush, H Jasper Senescent cells perturb intestinal stem cell differentiation through Ptk7 induced noncanonical Wnt and YAP signaling Nature Communications, 2023-01-11;14(1):156. 2023-01-11 [PMID: 36631445] (ELISA Capture, Mouse)

Bioinformatics

• Uniprot:
  • P27467()