Recombinant Human Frizzled-8 Fc Chimera Protein, CF

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

Recombinant Human Frizzled-8 Fc Chimera Protein, CF Summary

• Details of Functionality: Measured by its ability to inhibit Wnt-3a-induced alkaline phosphatase production by MC3T3‑E1 mouse preosteoblast cells. The ED₅₀ for this effect is 15-75 ng/mL in the presence of 10 ng/mL of Recombinant Mouse Wnt‑3a (Catalog # 1324-WN) Measured by its binding ability in a functional ELISA. In a 100 µL reaction mixture containing biotinylated recombinant mouse (rm) Wnt-3a or rmWnt-5a at 50-100 ng/mL and Recombinant Human (rh) Frizzled-8 Fc Chimera dilutions at 0.1-8,000 ng/mL, the concentration of rhFrizzled-8 Fc Chimera that produces 50% of the optimal binding response is found to be approximately 7-28 ng/mL.
  Optimal concentrations should be determined by each laboratory for each application.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Frizzled-8 protein
  

  Human Frizzled-8 (Ala25-Pro172) Accession # Q9H461	IEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: Q9H461
• N-terminal Sequence: Ala25 & Ala28
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• Gene: FZD8
• 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
• Theoretical MW: 43.3 kDa (monomer).
  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: 55-65 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.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 500 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 Human Frizzled-8 Fc Chimera Protein, CF

• frizzled (Drosophila) homolog 8
• frizzled 8
• frizzled family receptor 8
• frizzled homolog 8 (Drosophila)
• Frizzled8
• Frizzled-8
• FZ-8
• FZD8
• hFZ8

Background

Frizzled-8 is one of at least ten seven-transmembrane (7TM) glycoproteins of the Frizzled family of Wnt receptors (1-2). Frizzled proteins are thought to be G-protein-coupled (1). Wnt engagement, with low density lipoprotein receptor-related proteins LRP-5 or LRP-6 acting as co-receptors, stabilizes beta -catenin and promotes gene transcription that is important in development and tissue maintenance (1). Human Frizzled-8 cDNA encodes 694 amino acids (aa) including a 27 aa signal peptide, a 247 aa extracellular domain (ECD), the 7TM region (aa 276-605), and a C-terminal cytoplasmic domain with a PDZ binding motif (aa 606-694) (2). The ECD includes a cysteine-rich region (CRD, aa 30-151) that binds Wnts and is highly conserved among Frizzleds, and a linker region. Within aa 28-172, human Frizzled-8 shares 99%, 99%, 90% and 85% aa identity with mouse, rat, Xenopus and zebrafish Frizzled-8, respectively. It also shares 82% aa identity with human Frizzled-5. Frizzleds can form homodimers or selective hetero-oligomers with other family members, which can involve the TM regions and possibly the CRD (1, 3). During mouse development, Frizzled-8 is expressed in tissues that are important for organizing the anterior-posterior axis (4). Interactions of Frizzled-8 with several non-Wnt ligands have been identified. Interaction with Frizzled-8 and LRP-6 allows R-spondins to activate beta -catenin signaling pathways, while interaction with IGFBP-4 (insulin-like growth factor binding protein (4) or CTGF (connective tissue growth factor) inhibits Wnt signaling (5-7). These ligands bind the extracellular CRD of Frizzled-8, blocking Wnt binding. The recombinant Frizzled-8 CRD has also been used to block Wnt signaling and inhibit growth of teratocarcinomas (5-10).

1. Schulte, G. and V. Bryja (2007) Trends Pharmacol. Sci. 28:518.
2. Saitoh, T. et al. (2001) Int. J. Oncol. 18:991.
3. Dann, C.E. et al. (2001) Nature 412:86.
4. Lu C.C. et al. (2004) Gene Expr. Patterns 4:569.
5. Nam, J-S. et al. (2006) J. Biol. Chem. 281:13247.
6. Zhu, W. et al. (2008) Nature 454:345.
7. Mercurio, S. et al. (2004) Development 131:2137.
8. Chesnutt, C. et al. (2004) Dev. Biol. 274:334.
9. Yamashita, J.K. et al. (2005) FASEB J. 19:1534.
10. DeAlmeida, V.I. et al. (2007) Cancer Res. 67:5371.

Publications for Frizzled-8 (6129-FZ)(4)

Publications using 6129-FZ

• R Chidiac, M Abedin, G Macleod, A Yang, PE Thibeault, LL Blazer, JJ Adams, L Zhang, H Roehrich, HN Jo, S Seshagiri, SS Sidhu, HJ Junge, S Angers A Norrin/Wnt surrogate antibody stimulates endothelial cell barrier function and rescues retinopathy Embo Molecular Medicine, 2021-06-09;0(0):e13977. 2021-06-09 [PMID: 34105895] (Bioassay, Human)
• K Saito-Diaz, H Benchabane, A Tiwari, A Tian, B Li, JJ Thompson, AS Hyde, LM Sawyer, JN Jodoin, E Santos, LA Lee, RJ Coffey, RD Beauchamp, CS Williams, AK Kenworthy, DJ Robbins, Y Ahmed, E Lee APC Inhibits Ligand-Independent Wnt Signaling by the Clathrin Endocytic Pathway Dev. Cell, 2018-03-12;44(5):566-581.e8. 2018-03-12 [PMID: 29533772] (Bioassay, Human)
• Novel Bi-specific Domain Antibody to LRP6 Inhibits Wnt and R-spondin Ligand-induced Wnt Signaling and Tumor Growth Mol Cancer Res, 2016-07-11;0(0):. 2016-07-11 [PMID: 27401612] (Bioassay, Human)
• Wilson TR, Fridlyand J, Yan Y, Penuel E, Burton L, Chan E, Peng J, Lin E, Wang Y, Sosman J, Ribas A, Li J, Moffat J, Sutherlin DP, Koeppen H, Merchant M, Neve R, Settleman J Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature, 2012-07-26;487(7408):505-9. 2012-07-26 [PMID: 22763448] (Bioassay, Human)

Bioinformatics

• Gene Symbol: FZD8
• Uniprot:
  • Q9H461()