Frizzled-8 Antibody (1022733) [Unconjugated]

Frizzled‑8 was detected in immersion fixed PC‑3 human prostate cancer cell line (positive staining) and U937 human histiocytic lymphoma cell line (negative control) using Mouse Anti-Human Frizzled‑8 Monoclonal Antibody (Catalog # MAB6129) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Hu
• Applications: ICC/IF
• Clone: 1022733
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Frizzled-8 Antibody (1022733) [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Frizzled-8
  Ala25-Pro172
  Accession # Q9H461
• Specificity: Detects human Frizzled-8 in direct ELISAs.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL

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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

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

Alternate Names for Frizzled-8 Antibody (1022733) [Unconjugated]

• 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • Q9H461()