Recombinant Human Frizzled-8 Fc Biotinylated Protein, CF

When Recombinant Human Wnt‑3a (Catalog # 5036-WN) isimmobilized at 1 µg/mL (100 µL/well), Biotinylated Recombinant Human Frizzled‑8Fc Chimera (Catalog # BT6129) binds with an ED50 of 0.5-3 μg/mL.

2 μg/lane of Biotinylated Recombinant Human Frizzled-8 FcChimera (Catalog # BT6129) was resolved with SDS-PAGE under reducing (R) andnon-reducing (NR) conditions and visualized by Coomassie® Blue staining,showing bands at 53-63 kDa and 110-130 kDa, respectively.

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

Recombinant Human Frizzled-8 Fc Biotinylated Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human Wnt‑3a (Catalog # 5036-WN) is immobilized at 1 µg/mL (100 µL/well), Biotinylated Recombinant Human Frizzled‑8 Fc Chimera (Catalog # BT6129) binds with an ED₅₀ of 0.5-3 μg/mL.
• 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
• Structure / Form: Disulfide-linked homodimer, Biotinylated via amines
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 43 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: 56-63 kDa, under 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 with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• 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 Biotinylated 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.

Bioinformatics

• Uniprot:
  • Q9H461()