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Your Position: Home > Immune Checkpoint Proteins > CD155 > Biotinylated Human CD155 / PVR Protein, Fc,Avitag

Biotinylated Human CD155 / PVR Protein, Fc,Avitag™

INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
PVR,FLJ25946,PVS,CD155,TAGE4,HVED,NECL5
Source
Biotinylated Human CD155, Fc,Avitag (CD5-H82F6) is expressed from human 293 cells (HEK293). It contains AA Trp 21 - Asn 343 (Accession # AAH15542).
Predicted N-terminus: Trp 21
Request for sequence
Molecular Characterization
This protein carries a human IgG1 Fc tag at the C-terminus, followed by a Avi tag (Avitag™).
The protein has a calculated MW of 63.5 kDa. As a result of glycosylation, the protein migrates as 75-100 kDa under reducing (R) condition, and 140-180 kDa under non-reducing (NR) condition (SDS-PAGE).
Biotinylation
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
Biotin:Protein Ratio
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

Endotoxin
Less than 1.0 EU per μg by the LAL method.
Purity
>95% as determined by SDS-PAGE.
Formulation
Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
1. 4-8^°C for 12 months in lyophilized state;
2. -70^°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE

Biotinylated Human CD155, Fc,Avitag on SDS-PAGE under reducing (R) and non-reducing (NR) conditions. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity-ELISA

Biotinylated Human CD155, Fc,AvitagBiotinylated Human CD155, Fc,Avitag (Cat. No. CD5-H82F6) ELISA bioactivity

Immobilized Human TIGIT, Fc Tag (Cat. No. TIT-H5254) at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD155, Fc,Avitag (Cat. No. CD5-H82F6) with a linear range of 0.5-16 ng/mL (QC tested).

Protocol

Immobilized Human DNAM-1, Fc Tag (Cat. No. DN1-H5257) at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD155, Fc,Avitag (Cat. No. CD5-H82F6) with a linear range of 0.02-0.312 μg/mL (Routinely tested).

Protocol

Serial dilutions of TIGIT antibody antibody (1:2 serial dilutions, from 20 μg/mL to 0.0097 μg/mL) were added into Human TIGIT, His Tag (Cat. No. TIT-H52H3): Biotinylated Human CD155, Fc,Avitag (Cat. No. CD5-H82F6) binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points before curve fitting.

Protocol

Bioactivity-FACS

FACS assay shows that recombinant Biotinylated FACS

FACS assay shows that Biotinylated Human CD155, Fc,Avitag (Cat. No. CD5-H82F6) can bind to 293T cell overexpressing human TIGIT. The concentration of CD155 is 1 μg/mL (Routinely tested).

Protocol

FACS analysis shows that the binding of Biotinylated Human CD155, Fc,Avitag (Cat. No. CD5-H82F6) to 293T overexpressing TIGIT was inhibited by increasing concentration of neutralizing Anti-Human TIGIT MAb. The concentration of CD155 used is 1 μg/mL. The IC50 is 0.201 μg/mL (Routinely tested).

Protocol

Background
CD155 is a Type I transmembrane glycoprotein in the immunoglobulin superfamily. Commonly known as Poliovirus Receptor (PVR) due to its involvement in the cellular poliovirus infection in primates, CD155's normal cellular function is in the establishment of intercellular adherens junctions between epithelial cells. CD155/PVR was originally isolated based on its ability to mediate polio virus attachment to host cells. The fulllength (or CD155 alpha isoform) is synthesized as a 417 amino acid (aa) precursor that contains a 20 aa signal sequence, a 323 aa extracellular region, a 24 aa TM segment and a 50 aa cytoplasmic tail. The extracellular region contains one N terminal V type and two C2 type Ig like domains. CD155 is a transmembrane protein with 3 extracellular immunoglobulin-like domains, D1-D3, where D1 is recognized by the virus. Low resolution structures of CD155 complexed with poliovirus have been obtained using electron microscopy while a high resolution structures of theectodomain D1 and D2 of CD155 were solved by x-ray crystallography.
References
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.