Recombinant Human 5T4 Fc Chimera Avi-tag Protein, CF

Catalog #: AVI4975 Datasheet / COA / SDS

Biotinylated

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AVI4975-050

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

Recombinant Human 5T4 Fc Chimera Avi-tag Protein, CF Summary

Learn more about Avi-tag Biotinylated Proteins

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 Human 5T4 Antibody (Catalog # MAB49751) is immobilized at 0.5 µg/mL (100 µL/well), Biotinylated Recombinant Human 5T4 Fc Chimera Avi-tag binds with an ED₅₀ of 0.45-3.60 ng/mL. Measured by its binding ability in a functional ELISA. When Recombinant Human Frizzled-8 Fc Chimera (Catalog # 6129-FZ) is immobilized at 5 µg/mL (100 µL/well), Biotinylated Recombinant Human 5T4 Fc Chimera Avi-tag (Catalog # AVI4975) binds with an ED₅₀ of 5.00-40.0 µg/mL.

Source

Human embryonic kidney cell, HEK293-derived human 5T4 protein

Human 5T4 (Ser31-Ser355) Accession # Q13641.1	IEGRMD	Human IgG₁ (Pro100-Lys330)	Avi-tag
N-terminus			C-terminus

Accession #

Q13641.1

N-terminal Sequence
Analysis

Ser31

Structure / Form

Disulfide-linked homodimer, biotinylated via Avi-tag

Predicted Molecular Mass

64 kDa

SDS-PAGE

90-105 kDa, under reducing conditions

Product Datasheets

AVI4975

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.

AVI4975

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 Human 5T4 Antibody (MAB49751) is immobilized at 0.5 µg/mL (100 µL/well), Biotinylated Recombinant Human 5T4 Fc Chimera Avi-tag (Catalog # AVI4975) binds with an ED₅₀ of 0.45-3.60 ng/mL.

SDS-PAGE

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2 μg/lane of Biotinylated Recombinant Human 5T4 Fc Chimera Avi-tag (Catalog # AVI4975) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 90-105 kDa and 180-210 kDa, respectively.

Reconstitution Calculator

Background: 5T4

5T4, also known as Trophoblast glycoprotein and Wnt-activated inhibitory factor 1 (WAIF1), is a single-pass transmembrane protein which may function as an inhibitor of Wnt/beta-catenin signaling (1). Mature 5T4 consists of an extracellular domain (ECD) with eight leucine-rich repeats thought to antagonize Wnt signaling, a single transmembrane segment, and a short cytoplasmic tail with a PDZ-binding motif (2). Within the ECD, human 5T4 shares 78% and 77% amino acid sequence identity with mouse and rat 5T4, respectively. 5T4 has limited expression in normal adult tissues and in some solid tumors, while expression is elevated in human carcinomas which have been correlated with poor survival and prognosis (3-5). It has been identified that 5T4 interacts with TIP-2/GIPC, a common interacting protein involved in cancer, through its PDZ domain (6). 5T4 inhibits Wnt/ beta -catenin signaling by binding to LRP6 and inhibits Wnt-induced LRP6 internalization into endocytic vesicles (1). Conversely, 5T4 can activate noncanonical Wnt signaling through DKK1 (1). In addition to LRP6, in-house data indicates 5T4 can also interact with Frizzled-8. Our Avi-tag Biotinylated 5T4 features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

References

Kagermeier-Schenk, B. et al. (2011) Dev. Cell. 21:1129.
Zhao, Y. et al. (2014) Structure. 22:612.
Starzynska, T. et al. (1994) Br. J. Cancer. 69:899.
Naganuma, H. et al. (2002) Anticancer Res. 22:1033.
Harris, J.L. et al. (2018) Int. J. Biochem. Cell Biol. 99:28.
Awan, A. et al. (2002) BBRC 290:1030.

Long Name

Oncofetal Antigen/Trophoblast Glycoprotein

Entrez Gene IDs

7162 (Human); 21983 (Mouse); 83684 (Rat); 102132149 (Cynomolgus Monkey)

Alternate Names

5T4 oncofetal trophoblast glycoprotein; 5T4 oncotrophoblast glycoprotein; 5T4; 5T4AG; 5T4-AG; 5T4M6P1,5T4 oncofetal antigen; M6P1; TPBG; trophoblast glycoprotein

Related Research Areas

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