Human TIGIT Protein, Mouse IgG2a Fc Tag, low endotoxin

INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
TIGIT,VSIG9,VSTM3
Source
Human TIGIT, mouse IgG2a Fc tag (TIT-H5253) is expressed from human 293 cells (HEK293). It contains AA Met 22 - Pro 141 (Accession # AAI01290).
Predicted N-terminus: Met 22
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Molecular Characterization
This protein carries a mouse IgG2a Fc tag at the C-terminus.
The protein has a calculated MW of 40.1 kDa. The protein migrates as 42-50 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Endotoxin
Less than 0.1 EU per μg by the LAL method.
Purity
>95% as determined by SDS-PAGE.

Formulation
Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 100 mM Glycine, pH7.5. Normally trehalose is added as protectant before lyophilization.
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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 is 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, mouse IgG2a 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 95%.

Bioactivity

Immobilized Human CD155, Fc Tag (Cat. No. CD5-H5251) at 5μg/mL (100 μL/well) can bind Human TIGIT, mouse IgG2a Fc tag (Cat. No. TIT-H5253) with a linear range of 3.9-62.5 ng/mL (QC tested).

Request for protocol

Background
TIGIT (also called T cell immunoreceptor with Ig and ITIM domains) is one newly discovered immune receptor on some percentage of T cells and Natural Killer Cells(NK). It is also identified as WUCAM and Vstm3. TIGIT could bind to CD155(PVR) on dendritic cells(DCs), macrophages, etc. with high affinity, and also to CD112(PVRL2) with lower affinity. TIGIT expressing CD8+ T cells has been shown to be expanded and associated with clinical markers of HIV disease progression in a diverse group of HIV infected individuals. TIGIT and PD-1 has been shown to be over expressed on tumor antigen-sepcific (TA-specific) CD8+ T cells and CD8+ tumor infiltrating lymphocytes (TILs) from individuals with melanoma. Blockade of TIGIT and PD-1 led to increased cell proliferation, cytokine production, and degranulation of TA-specific CD8+ T cells and TIL CD8+ T cells.
References
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