Recombinant Mouse Wnt-9a Protein

Recombinant Mouse Wnt-9a (Catalog # 8148-WN) activates TCF reporter activity in HEK293 human embryonic kidney cells expressing human Frizzled-4 and human LRP-5. The ED50 for this effect is 8-40 ng/mL.

• Reactivity: Mu
• Applications: Bioactivity

Recombinant Mouse Wnt-9a Protein Summary

• Details of Functionality: Measured by its ability to activate Wnt induced TCF reporter activity in HEK293 human embryonic kidney cells expressing human Frizzled-4 and human LRP-5. The ED₅₀ for this effect is 8-40 ng/mL. Measured in a cell proliferation/survival assay using C3H10T1/2 mouse embryonic fibroblast cells. The ED₅₀ for this effect is 60-300 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived mouse Wnt-9a protein
  Tyr30-Gly365
• Accession #: Q8R5M2
• N-terminal Sequence: Tyr30
• Protein/Peptide Type: Recombinant Proteins
• Gene: Wnt9a
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 37 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: 36-43 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, EDTA and CHAPS with BSA as a carrier protein.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 250 μg/mL in 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 Mouse Wnt-9a Protein

• MGC138165
• MGC141991
• Protein Wnt-14
• wingless-type MMTV integration site family, member 14
• wingless-type MMTV integration site family, member 9A
• Wnt-14
• WNT14protein Wnt-9a
• Wnt9a
• Wnt-9a

Background

Wnt-9a (Wingless integration region 9a; also Wnt-14) is a cysteine-rich member of the Wnt family of secreted glycoproteins (1-4).  This family is rather large at 19 members, and although it is most often associated with embryogenesis, the Wnt family is now known to be also important in a number of related processes, including angiogenesis, wound healing, tissue remodeling, and cancer (2, 5-8). Wnt-9a represents a paralog (common ancestor; different function) to Wnt-9b/Wnt-14b which, in the mouse, are found on the same chromosome (2, 4). The mouse Wnt-9a precursor is 365 amino acids (aa) in length. It contains a 29 aa signal sequence coupled to a 336 aa mature region (aa 30-365) that is characterized by the presence of 24 Cys residues, two palmitoylation sites (Cys93 and Ser221), and one presumably utilized glycosylation site at Asn103 (4, 9). The glycosylation and palmitoylation appear essential for secretion and activity. Based on studies with other Wnts, the palmitic acid at Cys93 is essential for frizzled/FZD4, 7 and 9, WIF-1 activation, while Ser221 is necessary for secretion (10). Although the exact mechanism(s) for secretion is unclear, it would appear that Wnt-9a initially binds to a select GPCR (Evi) in the Golgi. As a complex, it is transported to the plasma membrane in where it is inserted, internalized, and budded-off in a microvesicle that returns to the cell membrane for release as an exosome (11). Over the mature region, mouse Wnt-9a is identical in aa sequence to rat Wnt-9a and shares 98% aa sequence identity with human Wnt-9a (12). With respect to its paralog (Wnt-9b), mouse Wnt-9a and -9b share only 65% aa sequence identity in the mature region (4, 13). Cells known to express Wnt-9a, either inducibly or constitutively, include skeletal muscle (14, 15), limb-bud mesenchyme (16), mammary epithelium (17, 18), embryonic hepatic stellate/Ito and sinusoidal endothelial cells (19), and cranial osteoblasts (20). There are an extraordinary number of receptors/binding proteins that have been identified for members of the Wnt family (1, 21, 22). To date, Wnt-9a has been reported to bind to Fzd4, 7 and 9 (19), WIF-1 (16), and MuSK in conjunction with LRP4 (15). Functionally, Wnt-9a is best known for its participation in joint formation (but not initiation). During the formation of cartilaginous tissue, Wnt-9a contributes to the process of cavitation where chondrogenic tissue undergoes apoptosis and remodeling to create a functional space (interzone; precursor to a joint) (23). Elsewhere in the embryo, Wnt-9a is posited to induce hepatocyte proliferation and promote the generation of glycogen through enzyme activity regulation (19). In addition, Wnt-9a is reported to induce acetylcholine receptor clustering in newly formed myotubes (15).

1. Kikuchi, A. et al. (2007) Cell. Signal. 19:659.
2. Katoh, M. (2007) Stem Cell Rev. 3:30.
3. Katoh, M. (2002) Int. J. Oncol. 21:1269.
4. Qian, J. et al. (2003) Genomics 81:34.
5. Miller, J. (2002) Genome Biol. 3:Reviews3001.
6. Newman, A.C. & C.W. Hughes (2012) Vasc. Cell 4:13.
7. Miao, C. et al. (2013) Cell. Signal. 25:2069.
8. Bielefeld, K.A. et al. (2013) Cell. Mol. Life Sci. 70:2059.
9. SwissProt #:Q8R5M2.
10. Ke, J. et al. (2013) Curr. Opin. Lipidol. 24:129.
11. Gross, J.C. et al. (2013) Nat. Cell Biol.  14:1036.
12. Katoh, Y. & M. Katoh (2005) Oncol. Rep. 13:989.
13. Kirikoshi, H. et al. (2001) Int. J. Oncol. 19:947.
14. Saitoh, T. et al. (2001) Biochem. Biophys. Res. Commun. 284:1168.
15. Zhang, B. et al. (2012) Mol. Brain 5:7.
16. Surmann-Schmitt, C. et al. (2009) J. Cell Sci. 122:3627.
17. Benhaj, K. et al. (2006) Oncol. Rep. 15:701.
18. Witte, F. et al. (2009) Gene Exp. Patterns 9:215.
19. Matsumoto, K. et al. (2008) Dev. Biol. 319:234.
20. Zhou, H. et al. (2009) Development 136:427.
21. Hendrickx, M & L. Leyns (2008) Develop. Growth Differ. 50:229.
22. Kawano, Y & R. Kypta (2003) J. Cell Sci. 116:2627.
23. Spater, D. et al. (2006) Development 133:3039.

Publications for Wnt-9a (8148-WN)(1)

Publication using 8148-WN

• Congwei Luo, Shan Zhou, Zhanmei Zhou, Yahong Liu, Li Yang, Jiafeng Liu et al. Wnt9a Promotes Renal Fibrosis by Accelerating Cellular Senescence in Tubular Epithelial Cells Journal of the American Society of Nephrology 2018-04-01 [PMID: 29440280] (Bioassay, Human)

Bioinformatics

• Gene Symbol: Wnt9a
• Uniprot:
  • Q8R5M2()