Human TIGIT Protein, Fc Tag

INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
TIGIT,VSIG9,VSTM3
Source
Human TIGIT, Fc Tag (TIT-H5254) is expressed from human 293 cells (HEK293). It contains AA Met 22 - Pro 141 (Accession # Q495A1 -1).
Predicted N-terminus: Met 22
Request for sequence
Molecular Characterization
This protein carries a human IgG1 Fc tag at the C-terminus.
The protein has a calculated MW of 39.7 kDa. The protein migrates as 42-45 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Endotoxin
Less than 1.0 EU per μg by the LAL method.
Purity
>90% as determined by SDS-PAGE.

Formulation
Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 100 mM Glycine, 25 mM Arginine, 10 mM 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

Human TIGIT, Fc Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity-ELISA

Human TIGIT, Fc TagHuman TIGIT, Fc Tag (Cat. No. TIT-H5254) ELISA bioactivity

Immobilized Human CD155, mouse IgG2a Fc tag, low endotoxin (Cat. No. CD5-H5254) at 5 μg/mL (100 μL/well) can bind Human TIGIT, Fc Tag (Cat. No. TIT-H5254) with a linear range of 0.6-5 ng/mL (QC tested).

Request for protocol

Immobilized Human TIGIT, Fc Tag (Cat. No. TIT-H5254) at 10 μg/mL (100 μL/well) can bind Biotinylated Human Nectin-2, Fc Tag (Cat. No. CD2-H82F8) with a linear range of 0.313-10 μg/mL (Routinely tested).

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FACS assay shows that recombinant Biotinylated FACS

FACS assay shows that recombinant Human TIGIT Protein, Fc Tag (Cat. No. TIT-H5254) can bind to 293T cell overexpressing human CD155. The concentration of TIGIT used is 0.1 μg/ml (Routinely tested).

Request for protocol

Background
T-cell immunoreceptor with Ig and ITIM domains (TIGIT) is also known as V-set and immunoglobulin domain-containing protein 9 (VSIG9), V-set and transmembrane domain-containing protein 3 (VSTM3),which belongs to single-pass type I membrane protein containing an immunoglobulin variable domain, a transmembrane domain and an immunoreceptor tyrosine-based inhibitory motif (ITIM). TIGIT is expressed at low levels on peripheral memory and regulatory CD4+ T-cells and NK cells and is up-regulated following activation of these cells (at protein level). TIGIT binds with high affinity to the poliovirus receptor (PVR) which causes increased secretion of IL10 and decreased secretion of IL12B and suppresses T-cell activation by promoting the generation of mature immunoregulatory dendritic cells.
References
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