Cynomolgus Monkey TIGIT Antibody

Catalog #: MAB10532 Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB10532-100
MAB10532-SP

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

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Cynomolgus Monkey TIGIT Antibody Summary

Species Reactivity

Cynomolgus Monkey

Specificity

Detects Cynomolgus Monkey TIGIT in direct ELISAs.

Source

Recombinant Monoclonal Rabbit IgG Clone # 2629A

Purification

Protein A or G purified from cell culture supernatant

Immunogen

Human embryonic kidney cell HEK293-derived Cynomolgus monkey TIGIT
Met22-Pro142
Accession # XP_00554815

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

HEK293 Human Cell Line transfected with Cynomolgus Monkey TIGIT and eGFP

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.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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

Blockade of Receptor-ligand Interaction

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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