Mouse TIGIT Antibody

Catalog #: AF7267 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7267
AF7267-SP

Detection of TIGIT in Mouse Splenocytes by Flow Cytometry.

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Mouse TIGIT Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse TIGIT in direct ELISAs. In direct ELISAs, less than 5% cross-reactivity with recombinant human TIGIT is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse TIGIT
Gly26-Thr143
Accession # NP_001139797

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

2.5 µg/10⁶ cells

See below

Immunohistochemistry

3-15 µg/mL

Immersion fixed paraffin-embedded sections of mouse spleen

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 Mouse Splenocytes by Flow Cytometry. 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).

Immunohistochemistry

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TIGIT in Mouse Spleen. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

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 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.

Long Name

T Cell Immunoreceptor with Ig and ITIM Domains

Entrez Gene IDs

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

Alternate Names

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