TIGIT Antibody [Unconjugated]

Mouse splenocytes treated with 50 ng/mL PMA and 500 ng/mL Calcium ionomycin for 24 hours were stained with Sheep Anti-Mouse TIGIT Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7267) followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127) and Rat Anti‑Mouse CD4 PE‑conjugated Monoclonal Antibody (Catalog # FAB554P). Quadrant markers were set based on control antibody staining (Catalog # 5-001-A).

TIGIT was detected in immersion fixed paraffin-embedded sections of mouse spleen using Sheep Anti-Mouse TIGIT Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7267) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Mu
• Applications: Flow, IHC, CyTOF-ready
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

TIGIT Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse TIGIT
  Gly26-Thr143
  Accession # NP_001139797
• Specificity: Detects mouse TIGIT in direct ELISAs. In direct ELISAs, less than 5% cross-reactivity with recombinant human TIGIT is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: Tigit
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunohistochemistry 3-15 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

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

Alternate Names for TIGIT Antibody [Unconjugated]

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

Background

TIGIT (T cell Immunoreceptor with Ig and ITIM domains; also Vstm3 and Vsig9) is a 30-34 kDa (in human) member of the CD28 family, Ig superfamily of molecules. It is expressed by NK cells and multiple subsets of mature T cells, and binds to PVR/CD155 and PVR2/CD112 that appear on dendritic cells (DC) and endothelium. Along with CD226, the TIGIT:CD226/DNAM1 and PVR:PVR2 pairings appear to form a network that parallels the well-characterized B7-1:B7-2 and CD28:CTLA4 system. Binding of TIGIT by DC induces DC IL-10 release and inhibits IL-12 production. Ligation of TIGIT on T cells dampens TCR-mediated activation, while NK cell TIGIT ligation blocks NK cell cytotoxicity. Mature mouse TIGIT is a type I transmembrane protein 215 amino acids (aa) in length. It contains a 116 aa extracellular region (aa 26-141) with a V-type Ig-like domain (aa 27-125), and a 79 aa cytoplasmic domain with one ITIM motif. There is one isoform variant that is quite unusual and shows an addition of nine amino acids spread over three insertion sites (SwissProt #:P86176). Mouse and human TIGIT are highly divergent, and over aa 26-143, mouse TIGIT shares only 68% aa sequence identity with human TIGIT.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: Tigit
• Uniprot:
  • NP_001139797()