Anti-Mouse TIGIT [Clone 1G9] — Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse TIGIT [Clone 1G9] — Purified in vivo PLATINUM™ Functional Grade

Product No.: T736

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Clone
1G9
Target
TIGIT
Formats AvailableView All
Product Type
Hybridoma Monoclonal Antibody
Alternate Names
Vstm3, VSIG9
Isotype
Mouse IgG1 κ
Applications
ELISA
,
FA
,
FC

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Mouse
Recommended Dilution Buffer
Immunogen
Recombinant murine TIGIT tetramers.
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
≤ 0.5 EU/mg as determined by the LAL method
Purity
≥98% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
State of Matter
Liquid
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM<sup>TM</sup> antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Regulatory Status
Research Use Only
Country of Origin
USA
Shipping
2 – 8° C Wet Ice
Additional Applications Reported In Literature ?
ELISA,
FA,
FC
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
Clone 1G9 activity is directed against mouse TIGIT.
Background
TIGIT is an immunoreceptor that inhibits multiple immune cell responses, including T cell priming by dendritic cells, tumor cell killing by NK cells and cytotoxic T cells, and also enhances the immune suppressive activity of regulatory T cells1. TIGIT is a novel member of the Ig-superfamily distantly related to Nectins and Necls that aligns with the distal Ig-V-type domains of Nectin1-4, poliovirus receptor (PVR; CD155), DNAM-1 (CD226), and TACTILE (CD96)2. TIGIT is an attractive target for cancer therapy due to its role as an immune checkpoint1,3. Immunotherapy targeting TIGIT and the PD-1/PD-L1 pathway is capable of tumor suppression.

1G9 was generated by immunizing TIGIT-/- mice with recombinant mouse TIGIT tetramers3. Draining lymph nodes were collected and fused with Sp2/0-Ag14. Supernatants were screened for specific binding by anti-TIGIT ELISA and flow cytometry. Hybridomas that showed TIGIT-specific binding were expanded and subcloned and single colonies sorted by flow cytometry. Comparative immunofluorescence staining of activated primary TIGIT-expressing wildtype T cells and TIGIT-/- T cells was performed to confirm specificity. 1G9 was found to fully block TIGIT binding to CD155, a high-affinity TIGIT ligand. However, 1G9 does not deplete TIGIT+ cells in vivo under steady-state conditions. Additionally, 1G9 does not affect T cell proliferation in vitro. 1G9 has agonistic anti-TIGIT activity in vivo, leading to a reduction in T cell expansion and pro-inflammatory cytokine production, and is also able to reduce experimental autoimmune encephalomyelitis (EAE) severity in mice.
Antigen Distribution
TIGIT is expressed on NK cells, activated T cells, memory T cells, and a subset of regulatory T cells.
Ligand/Receptor
CD155 (PVR) and CD112 (PVRL2)
NCBI Gene Bank ID
UniProt.org
Research Area
Cell Biology
.
Immunology

Leinco Antibody Advisor

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In Vivo Applications of Clone 1G9 in Mice

Clone 1G9 is a mouse monoclonal antibody (IgG1, κ) that specifically recognizes mouse TIGIT (T cell immunoreceptor with Ig and ITIM domains). Its in vivo use is primarily as an agonistic antibody to modulate immune responses mediated by TIGIT, an inhibitory immune checkpoint receptor expressed on T cells, including regulatory T cells (Tregs) and subsets of activated and memory T cells.

Key In Vivo Applications

  • Suppression of T Cell-Driven Inflammation: Clone 1G9 is used in vivo to suppress T cell-driven inflammation in mice. By delivering agonistic signals through TIGIT, it reduces T cell expansion and the production of pro-inflammatory cytokines, thereby attenuating autoimmune and inflammatory responses. This makes it a valuable tool for studying the role of TIGIT in autoimmune diseases and immune regulation.
  • Investigation of Immune Checkpoint Modulation: As an immune checkpoint molecule, TIGIT negatively regulates T cell and NK cell activation. In vivo administration of 1G9 allows researchers to probe the consequences of TIGIT pathway engagement, including effects on immune tolerance, cytokine profiles, and the balance between immune activation and suppression.
  • Blockade of TIGIT/CD155 Interaction: In vitro, 1G9 fully blocks the binding of TIGIT to its high-affinity ligand CD155. While in vivo experiments confirm that 1G9 does not deplete TIGIT+ cells under steady-state conditions, its primary effect is functional blockade of the TIGIT pathway rather than cell depletion.
  • Flow Cytometry and Phenotyping: Although primarily an in vivo tool, 1G9 is also used for flow cytometry to identify and characterize TIGIT-expressing cell populations in mice. However, its most distinctive utility is in modulating immune responses in living animals.

Experimental Impact

  • Reduced T Cell Expansion: In vivo, 1G9 treatment leads to decreased T cell proliferation, highlighting TIGIT’s role in restraining excessive immune activation.
  • Diminished Pro-Inflammatory Cytokine Production: The antibody reduces the secretion of pro-inflammatory cytokines, consistent with TIGIT’s immunosuppressive function.
  • No Cell Depletion: Unlike some therapeutic antibodies, 1G9 does not cause depletion of TIGIT-expressing cells, allowing researchers to study the functional consequences of pathway engagement without confounding effects of cell loss.

Summary Table

Application AreaEffect of Clone 1G9 In VivoKey Outcome
Autoimmune/Inflammatory ModelsAgonistic engagement of TIGITSuppressed T cell expansion, reduced inflammation
Immune Checkpoint StudiesFunctional blockade of TIGIT/CD155Altered immune tolerance, cytokine profiles
Phenotyping (supporting)Detection of TIGIT+ cells (flow cytometry)Identification of TIGIT-expressing subsets

Conclusion

Clone 1G9 is principally employed in mice to agonize TIGIT signaling in vivo, thereby suppressing T cell-driven inflammation and modulating immune checkpoint activity without depleting TIGIT+ cells. This makes it a critical reagent for investigating the immunoregulatory functions of TIGIT in autoimmune, inflammatory, and tolerance models.

Clone 1G9 is most commonly used as an anti-mouse TIGIT monoclonal antibody, targeting the immunoregulatory protein TIGIT on T and NK cells. In the literature, several other antibodies or proteins are frequently used alongside 1G9, largely because TIGIT biology is intimately connected to related checkpoint proteins and their ligands in immunological research, particularly cancer and autoimmunity studies.

The most commonly used antibodies/proteins with 1G9 include:

  • Anti-PD-1 (Programmed cell death protein 1) and anti-PD-L1 (Programmed death ligand 1):Researchers often explore immune checkpoint blockade by combining TIGIT and PD-1/PD-L1 antagonists to examine synergy in tumor suppression or immune modulation.

  • Anti-CD226 (DNAM-1):Because DNAM-1 (CD226) is a co-activating receptor that shares ligands (CD112 and CD155) with TIGIT and exhibits counter-regulatory function, studies often measure or block DNAM-1 signaling in parallel with TIGIT to dissect their distinct effects on immune cell activation.

  • Ligands: CD155 (PVR) and CD112 (PVRL2):These are the principal ligands for TIGIT and DNAM-1; detection or functional blocking antibodies/proteins against these ligands are frequently used to assess the consequences of interfering with the TIGIT-CD155 axis.

  • Anti-CD96 (TACTILE):Like TIGIT, CD96 interacts with CD155 and is another checkpoint under investigation; studies may use anti-CD96 in multiplex approaches with 1G9 when mapping regulatory pathways.

Summary Table: Common Antibodies/Proteins Used with 1G9

Partner Antibody/ProteinTargetReason for Use With 1G9
Anti-PD-1/PD-L1Immune checkpoint proteinsEvaluate synergistic checkpoint blockade
Anti-CD226 (DNAM-1)Activating co-receptorStudy interplay with TIGIT/CD155 pathway
Anti-CD155 (PVR), CD112Ligands for TIGIT, DNAM-1, CD96Functional pathway dissection
Anti-CD96Alternate checkpoint on same pathwayExamine redundant/parallel suppressive roles

Additional context:

  • Flow cytometry panels and functional assays using 1G9 will typically include markers for T cells (CD3, CD4, CD8), NK cells (NK1.1, CD49b), and regulatory T cells (FoxP3, CD25) to define affected populations.
  • In cancer and autoimmunity research, anti-TIGIT (1G9) is almost always used in the context of multicolor immunophenotyping or functional checkpoint blockade assays that require comprehensive immune monitoring.

According to reagent suppliers and published reviews on TIGIT biology, these combinations are standard in the literature using the 1G9 clone antibody.

Key Findings of Clone 1G9 Anti-TIGIT Antibody in Scientific Literature

Agonistic Activity and T Cell Modulation

  • Clone 1G9 is an agonistic anti-mouse TIGIT antibody that fully blocks the binding of TIGIT to its high-affinity ligand CD155 (PVR), demonstrating high specificity for mouse TIGIT.
  • In vivo, clone 1G9 exhibits agonistic activity, leading to reduced T cell expansion and decreased production of pro-inflammatory cytokines (e.g., IFN-γ and IL-17), particularly in models of antigen-specific T cell recall responses.
  • While clone 1G9 does not directly affect T cell proliferation in vitro, it significantly reduces antigen-specific proliferation and cytokine secretion upon antigen re-stimulation in treated mice, though to a lesser extent than clone 4D4.
  • EAE Models: Treatment with clone 1G9 reduces disease severity in experimental autoimmune encephalomyelitis (EAE) models, correlating with decreased Th17 cell frequencies in the central nervous system (CNS).
  • No Cell Depletion: Clone 1G9 does not deplete TIGIT-expressing cells in vivo under steady-state conditions, distinguishing it from antibody-dependent cell depletion strategies.

Functional Specificity and Mechanism

  • High Specificity: Clone 1G9 was generated by immunizing TIGIT knockout mice with recombinant mouse TIGIT tetramers, followed by extensive screening for TIGIT-specific binding using ELISA and flow cytometry.
  • Mechanism: The agonistic activity of clone 1G9 in vivo likely results from stabilizing inhibitory TIGIT signaling, thus dampening T cell activation and inflammatory responses, rather than blocking TIGIT function (as seen with clone 1B4, which acts as a blocking antibody and exacerbates disease).
  • TIGIT as Immune Checkpoint: TIGIT is an immune checkpoint receptor expressed on NK cells, activated and memory T cells, and a subset of regulatory T cells. Targeting TIGIT, especially in combination with PD-1/PD-L1 blockade, is considered a promising strategy for immunotherapy.

Contrast with Other Clones

CloneActivity TypeEffect on T Cell ProliferationEffect on Cytokine SecretionImpact on EAE SeverityNotes
1G9AgonisticReduces (in vivo)Reduces IFN-γ, IL-17 (in vivo)Reduces severityNo cell depletion
4D4AgonisticReduces (dose-dependent)Reduces IFN-γ, IL-17Reduces severity (hamster)Less effective in chronic models (species?)
1B4BlockingIncreasesIncreases IFN-γ, IL-17Increases severityPhenocopies TIGIT knockout

Summary Table of Key Effects

  • Reduces T cell expansion and pro-inflammatory cytokine production in vivo.
  • Attenuates autoimmune disease severity (EAE) by reducing Th17 cell infiltration.
  • Does not affect T cell proliferation in vitro, nor deplete TIGIT+ cells in vivo.
  • Fully blocks TIGIT:CD155 interaction without cell depletion.

Conclusion

Clone 1G9 is a well-characterized, agonistic anti-mouse TIGIT monoclonal antibody that modulates immune responses by suppressing T cell activation and cytokine production in vivo, without depleting TIGIT-expressing cells. Its therapeutic potential is highlighted by reduced severity in autoimmune models, making it a valuable tool for studying TIGIT biology and for exploring immunotherapeutic strategies targeting this checkpoint.

Dosing regimens of the anti-mouse TIGIT antibody clone 1G9 vary depending on the mouse model and experimental context, with doses typically ranging from 200 μg to 400 μg per mouse, and the schedule tailored to disease kinetics and study aims.

  • In a sepsis model (cecal ligation and puncture, CLP), 1G9 was administered subcutaneously at 400 μg per mouse 1 hour after CLP, with a second injection 12 hours later.
  • In an experimental autoimmune encephalomyelitis (EAE) model (a T cell-driven autoimmunity model), 200 μg per mouse intraperitoneally was used, administered on days 0, 2, and 4 after immunization.
  • Commercial and scientific overviews confirm no fixed standardized dosing guide exists for clone 1G9 across all models; choices are adjusted according to disease kinetics and experimental aims.

Key points:

  • Dose: Common regimens range from 200 μg to 400 μg per mouse per dose.
  • Administration route: Both subcutaneous (s.c.) and intraperitoneal (i.p.) injections are used, depending on the study.
  • Schedule: Dosing can be once, 12 hours apart, or multiple times (e.g., every other day) based on disease model and experimental design.
  • Model-specific adjustments: Dosing and scheduling are adapted for acute (e.g., sepsis) and chronic (e.g., autoimmunity, tumor) models, reflecting differences in antibody clearance and disease progression rates.

No data in the provided sources indicate lower threshold doses (e.g., 5 μg) for 1G9; the lowest and most common effective doses reported are in the 200–400 μg range.

If your application or disease model differs—for example, in oncology or infection—dosing regimens might need further adjustment, usually within the published range and based on factors such as disease kinetics, antibody half-life, and the readout of interest.

References & Citations

1 Harjunpää H, Guillerey C. Clin Exp Immunol. 200(2):108-119. 2020.
2 Boles KS, Vermi W, Facchetti F, et al. Eur J Immunol. 39(3):695-703. 2009.
3 Dixon KO, Schorer M, Nevin J, et al. J Immunol. 200(8):3000-3007. 2018.
4 Chen Y, Huang H, Li Y, et al. Front Immunol. 13:832230. 2022.
5 Zhou XM, Li WQ, Wu YH, et al. Front Immunol. 9:2821. 2018.
6 Wu L, Mao L, Liu JF, et al. Cancer Immunol Res. 7(10):1700-1713. 2019.
7 Peng H, Li L, Zuo C, et al. Front Immunol. 13:1039226. 2022.
8 Stirm K, Leary P, Wüst D, et al. J Immunother Cancer. 11(2):e006263. 2023.
9 Schorer M, Rakebrandt N, Lambert K, et al. Nat Commun. 11(1):1288. 2020.
10 Freed-Pastor WA, Lambert LJ, Ely ZA, et al. Cancer Cell. 39(10):1342-1360.e14. 2021.
11 Ozmadenci D, Shankara Narayanan JS, et al. Proc Natl Acad Sci U S A. 119(17):e2117065119. 2022.
Indirect Elisa Protocol
FA
Flow Cytometry

Certificate of Analysis

Formats Available

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.