Recombinant Human TIGIT (A103) Fc Chimera Protein, CF

Recombinant Human TIGIT (A103) Fc Chimera (Catalog # 7898-TGB) has a molecular weight (MW) of 81.6 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

When Recombinant Human CD155/PVR (2530-CD) is immobilized at 2.5 µg/mL, Recombinant Human TIGIT (A103) Fc Chimera (Catalog # 7898-TGB) binds with an ED50 of 40-200 ng/mL.

Recombinant Human CD155/PVR Fc protein (9174-CD) was immobilized on a Biacore Sensor Chip CM5, and binding to Recombinant Human TIGIT (A103) Fc protein (Catalog # 7898-TGB) was measured at a concentration range between 0.531 nM and 544 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of KD=67.7 nM.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human TIGIT (A103) Fc Chimera Protein, CF Summary

• Additional Information: Analyzed by SEC-MALS
• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human CD155/PVR (Catalog # 2530-CD) is coated at 2.5 μg/mL (100 μL/well), the concentration of Recombinant Human TIGIT (A103) Fc Chimera that produces 50% optimal binding response is 40-200 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human TIGIT protein
  

  Human TIGIT (Met22-Pro141, A103) Accession # AAI01289	IEGRMD	Human IgG1 (Pro100-Lys330)
  N-terminus		C-terminus

• Accession #: AAI01289
• N-terminal Sequence: Met22
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 40 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 40-53 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, ≤ -20 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TIGIT (A103) Fc Chimera Protein, CF

• T cell immunoreceptor with Ig and ITIM domains
• TIGIT
• VSIG9
• VSTM3
• WUCAM

Background

TIGIT (T cell Immunoreceptor with Ig and ITIM domains), also called Vstm3 (V-set and transmembrane domain-containing 3), Vsig9 (V-set and Ig domain-containing 9) and WUCAM (Washington University cell adhesion molecule) is a 30-34 kDa type I transmembrane protein that is a member of the CD28 family within the Ig superfamily of proteins (1-4). Human TIGIT cDNA encodes 244 amino acids (aa) including a 21 aa signal sequence, a 120 aa extracellular region with a V-type Ig-like domain and two potential N-glycosylation site, a 21 aa transmembrane sequence, and an 82 aa cytoplasmic domain with an ITIM motif (5). A 170 aa variant diverges after aa 166 (5). Within the ECD, human TIGIT shares only 68-75% aa sequence identity with mouse, porcine, canine, equine and bovine TIGIT. TIGIT is expressed on NK cells and subsets of activated, memory and regulatory T cells, and particularly on follicular helper T cells within secondary lymphoid organs (1, 2, 6-8). It binds to CD155/PVR/Necl-5 and Nectin-2/CD112/PVRL2 that appear on dendritic cells (DC) and endothelium (1-3, 7). Binding of TIGIT by DC induces IL-10 release and inhibits IL-12 production (2). Ligation of TIGIT on T cells down‑regulates TCR-mediated activation and subsequent proliferation, while NK cell TIGIT ligation blocks NK cell cytotoxicity (6-8). Through CD155 and Nectin-2, which also interact with DNAM-1/CD226 and CD96/Tactile, TIGIT is part of an interacting network of Ig superfamily members that may augment or oppose each other (3, 4, 6, 7). In particular, TIGIT binding to CD155 can antagonize the effects of DNAM-1 (6, 7). Soluble TIGIT is able to compete with DNAM-1 for CD155 binding and attenuates T cell responses, while mice lacking TIGIT show increased T cell responses and susceptibility to autoimmune challenges (2, 3, 8).

1. Boles, K.S. et al. (2009) Eur. J. Immunol. 39:695.
2. Yu, X. et al. (2009) Nat. Immunol. 10:48.
3. Levin, S.D. et al. (2011) Eur. J. Immunol. 41:902.
4. Xu, Z. et al. (2010) Cell. Mol. Immunol. 7:11.
5. SwissProt Accession # Q495A1.
6. Seth, S. et al. (2009) Eur. J. Immunol. 39:3160.
7. Stanietsky, N. et al. (2009) Proc. Natl. Acad. Sci. USA 106:17858.
8. Joller, N. et al. (2011) J. Immunol. 83:1338.

Publications for TIGIT (7898-TGB)(1)

Publication using 7898-TGB

• Wang F, Hou H, Wu S, Tang Q, Liu W, Huang M, Yin B, Huang J, Mao L, Lu Y, Sun Z TIGIT expression levels on human NK cells correlate with functional heterogeneity among healthy individuals. Eur J Immunol, 2015-09-07;45(10):2886-97. 2015-09-07 [PMID: 26171588] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • AAI01289()