Recombinant Human 5T4 His-tag Avi-tag Protein, CF

Biotinylated
Catalog # Availability Size / Price Qty
AVI10290-100
Recombinant Human 5T4 His-tag Avi-tag Protein, CF Binding Activity
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Recombinant Human 5T4 His-tag Avi-tag 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 Human 5T4 Antibody (Catalog # MAB49751) is immobilized at 0.5 µg/mL (100 µL/well), Biotinylated Recombinant Human 5T4 His-tag Avi-tag (Catalog # AVI10290) binds with an ED50 of 10-80 ng/mL.
Source
Chinese Hamster Ovary cell line, CHO-derived human 5T4 protein
Ser31-Ser355, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Ser31
Structure / Form
Biotinylated via Avi-tag
Predicted Molecular Mass
38 kDa
SDS-PAGE
40-80 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.

AVI10290

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 400 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: No Stability Data for Carrier Free, Lyophilized

Data Images

Binding Activity Binding Activity View Larger

When Human 5T4 Antibody (Catalog # MAB49751) is immobilized at 0.5 μg/mL (100 μL/well), Biotinylated Recombinant Human 5T4 His-tag Avi-tag (Catalog # AVI10290) binds with an ED50 of 10-80 ng/mL.

SDS-PAGE SDS-PAGE View Larger

2 μg/lane of Recombinant Human 5T4 His-tag Avi-tag was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 40-80 kDa.

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: 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). Distinct regions on the 5T4 ectodomain which contains eight leucine-rich repeats are thought to antagonize Wnt signaling (2). 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). Human 5T4 is a 420 amino acid (aa) protein that includes a 31 aa signal peptide, a 324 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 44 aa cytoplasmic tail. Within the ECD, human 5T4 shares 78% and 77% aa sequence identity with mouse and rat 5T4, respectively. It has been identified that TIP-2/GIPC, a common interacting protein involved in cancer, interacts with 5T4 through its PDZ domain (6). 5T4 inhibits Wnt/ beta -catenin signaling by binding to LRP6 and inhibit 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.

References
  1. Kagermeier-Schenk, B. et al. (2011) Dev. Cell. 21:1129.
  2. Zhao, Y. et al. (2014) Structure. 22:612.
  3. Starzynska, T. et al. (1994) Br. J. Cancer. 69:899.
  4. Naganuma, H. et al. (2002) Anticancer Res. 22:1033.
  5. Harris, J.L. et al. (2018) Int. J. Biochem. Cell Biol. 99:28.
  6. 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

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