HEK293 human embryonic kidney cell line transfected with either (A) cynomolgus monkey TIGIT or (B) irrelevant protein, and eGFP, was stained with Rabbit anti-cynomolgus monkey TIGIT monoclonal antibody (Catalog # MAB10532) followed by Allophycocyanin-conjugated anti-Rabbit IgG Secondary Antibody (F0111). Quadrant markers were set based on control antibody staining (MAB1050, data not shown). Staining was performed using our Staining Membrane-Associated Proteins protocol.

TIGIT binding to CD155/PVR blocked by TIGIT Antibody.

In a functional ELISA, Recombinant Cynomolgus Monkey TIGIT (9380-TG) binds to immobilized Recombinant Human CD155/PVR (2530-CD) coated at 2.5 µg/mL (100 µL/well) in a dose-dependent manner (orange line). Binding is blocked (green line) by increasing concentrations of Rabbit Anti-Cynomolgus Monkey TIGIT Monoclonal Antibody (Catalog # MAB10532). At 70-350 ng/mL, this antibody will block 50% of the binding.

Applications for Cynomolgus Monkey TIGIT Antibody

Application

Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 70-350 ng/mL of this antibody will block 50% of the binding of 125 ng/mL of Recombinant Cynomolgus Monkey TIGIT Fc Chimera (Catalog # 9380-TG) to immobilized Recombinant Human CD155/PVR (Catalog # 2530-CD) coated at 2.5 ug/mL (100 µL/well). At 10 ug/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 Human Cell Line transfected with Cynomolgus Monkey TIGIT and eGFP

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: TIGIT

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. Cyno TIGIT shares 88.4% homology with human 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).

References

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

Long Name

T Cell Immunoreceptor with Ig and ITIM Domains

Alternate Names

VSIG9, VSTM3, WUCAM

Entrez Gene IDs

201633 (Human); 100043314 (Mouse); 363784 (Rat); 102121588 (Cynomolgus Monkey)

Gene Symbol

TIGIT

UniProt

XP_00554815

Additional TIGIT Products

Product Documents for Cynomolgus Monkey TIGIT Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for Cynomolgus Monkey TIGIT Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Cynomolgus Monkey TIGIT Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB10532

Key Product Details

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Label

Antibody Source

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Product Specifications

Immunogen

Specificity

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Host

Isotype

Scientific Data Images for Cynomolgus Monkey TIGIT Antibody

Detection of TIGIT in HEK293 Human Cell Line transfected with Cynomolgus Monkey TIGIT and eGFP by Flow Cytometry.