Biotinylated Human CD155, Fc,Avitag (CD5-H82F6) is expressed from human 293 cells (HEK293). It contains AA Trp 21 - Asn 343 (Accession # NP_006496.2).
Predicted N-terminus: Trp 21
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. The protein migrates as 80-106 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5. Normally trehalose is added as protectant before lyophilization.
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Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human CD155, Fc,Avitag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
The purity of Biotinylated Human CD155, Fc,Avitag (Cat. No. CD5-H82F6) was more than 85% and the molecular weight of this protein is around 165-185 kDa verified by SEC-MALS.
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 7-62 ng/mL (QC tested).
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).
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.
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).
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.