Recombinant Human Frizzled-8 Fc Biotinylated Protein, CF

Catalog # Availability Size / Price Qty
BT6129-050
Recombinant Human Frizzled-8 Fc Biotinylated Protein, CF Binding Activity
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Recombinant Human Frizzled-8 Fc Biotinylated Protein, CF Summary

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
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-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Ala25
Structure / Form
Disulfide-linked homodimer, Biotinylated via amines
Predicted Molecular Mass
43 kDa
SDS-PAGE
56-63 kDa, under reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

BT6129

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Data Images

Binding Activity Binding Activity View Larger

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.

SDS-PAGE SDS-PAGE View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Frizzled-8

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).

References
  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.
Entrez Gene IDs
8325 (Human); 14370 (Mouse)
Alternate Names
frizzled (Drosophila) homolog 8; frizzled 8; frizzled family receptor 8; frizzled homolog 8 (Drosophila); Frizzled8; Frizzled-8; FZ-8; FZD8; hFZ8

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