Recombinant Human Frizzled-8 Fc Biotinylated Protein, CF

Catalog #: BT6129 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
BT6129-050

Recombinant Human Frizzled-8 Fc Biotinylated Protein Binding Activity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Frizzled-8 Fc Biotinylated Protein, CF Summary

Product Specifications

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-terminus		C-terminus

Accession #

Q9H461

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

BT6129

Product Datasheet

COA

SDS

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.

Scientific Data

Binding Activity

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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.

SDS-PAGE

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

Reconstitution Calculator

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