Anti-Mouse CXCL9 (Clone MIG-2F5-5) – Purified in vivo PLATINUM™ Functional Grade

Clone MIG-2F5-5 is most commonly used in vivo in mice to neutralize the chemokine CXCL9 (MIG), making it a key tool for studying immune cell trafficking, T-cell recruitment, and the functional role of CXCL9 in various immunological processes and disease models.

Essential in vivo applications include:

• CXCL9 neutralization: The primary use of MIG-2F5-5 is to block CXCL9 activity in murine models, thereby inhibiting its ability to attract CXCR3-expressing T cells and NK cells to sites of inflammation, tumors, or infection.
• Tumor immunity research: MIG-2F5-5 has been used to dissect the contribution of CXCL9-mediated T-cell recruitment in cancer models, particularly to understand how manipulation of the tumor immune microenvironment affects tumor growth and response to therapies. For example, it has been used to investigate how neutralizing CXCL9 alters the response to IFNα2-based immunotherapies and impacts tumor growth and metastasis.
• Autoimmune and inflammatory disease studies: Because CXCL9 regulates immune cell migration and localization, in vivo blockade is often employed to clarify the chemokine’s role in the pathogenesis of autoimmune or inflammatory conditions such as colitis, arthritis, and allograft rejection.

Supporting details:

• MIG-2F5-5 is an Armenian hamster IgG monoclonal antibody specific to murine CXCL9.
• It is suitable for functional grade in vivo applications due to its high purity and low endotoxin levels.
• In addition to in vivo neutralization, it is reported to work in immunofluorescence and some flow cytometry applications, though its chief utility is blockade of CXCL9 function in live mice.

In summary, clone MIG-2F5-5 is widely employed in mouse models to study the effects of CXCL9, with core applications spanning tumor immunology and immune cell trafficking in disease contexts through in vivo chemokine neutralization.

The MIG-2F5-5 antibody, which targets CXCL9, is often used alongside other proteins and antibodies involved in similar biological pathways. Some of the commonly used antibodies or proteins include:

• CXCL10 and CXCL11: These chemokines, like CXCL9, interact with the CXCR3 receptor and are involved in directing the trafficking of immune cells, such as T cells and NK cells.
• CXCR3: This receptor is crucial for the activity of CXCL9, CXCL10, and CXCL11, as it mediates their chemotactic effects on T cells.
• IFN-γ (Interferon gamma): This cytokine is a potent inducer of CXCL9 expression in various cell types, including macrophages.
• TIM-3 and Galectin-9: These molecules are involved in immune checkpoint regulation and can interact with pathways that include CXCL9/CXCR3 signaling.

These proteins and antibodies are frequently used in studies related to immune cell trafficking, inflammation, autoimmune diseases, and cancer research.

The MIG-2F5.5 monoclonal antibody has generated significant research findings across multiple areas of immunology and cancer biology, particularly regarding the role of CXCL9 in immune responses and tumor immunology.

Cancer Immunotherapy and Immune Checkpoint Inhibitors

Research using MIG-2F5.5 has revealed important correlations in human cancer patients between IFN-I pathway activation, CXCL9 expression, and T cell expansion following PD-(L)1 immune checkpoint inhibitor immunotherapy. These findings have been particularly compelling in demonstrating CXCL9's critical role in anti-tumor immunity. The antibody has proven instrumental in elucidating how CXCL9 contributes to the effectiveness of cancer immunotherapy approaches.

Tumor Microenvironment and Chemokine Signaling

Studies utilizing this clone have uncovered intricate relationships between different chemokines in the tumor microenvironment. Research with ovarian cancer models showed that tumor cell-derived CCL5 drives CXCL9 expression in tumor-associated macrophages (TAMs), which in turn determines the tumor immunophenotype. When CCL5 was knocked down in tumor cells, there was significantly decreased CXCL9 expression in tumor-infiltrating CD11b+ cells, along with reduced expression of immune activation markers including CD8a, IFN-γ, and granzyme B. This loss of CXCL9 expression resulted in accelerated tumor growth and shorter survival, confirming that tumor-intrinsic CCL5 expression drives CXCL9 production in myeloid cells.

Cellular Expression and Function

The MIG-2F5.5 antibody has been extensively validated for detecting CXCL9 expression in various cell types, including macrophages, monocytes, endothelial cells, fibroblasts, and cancer cells responding to IFN-γ stimulation. Flow cytometric studies using this antibody have demonstrated robust CXCL9 expression in stimulated RAW264.7 macrophage cells and peritoneal exudate cells following IFN-γ and LPS treatment. These experiments confirmed that CXCL9 is primarily expressed intracellularly and can be detected using appropriate fixation and permeabilization protocols.

Immunological Characteristics

Research has established that CXCL9 functions as a T-cell chemoattractant and is unusual among CXC chemokines for being chemotactic for lymphocytes rather than neutrophils. This distinguishes it from most other CXC family members and highlights its specific role in lymphocyte recruitment to sites of inflammation and tumor tissues.

Dosing regimens for clone MIG-2F5-5 (anti-mouse CXCL9) vary depending on the experimental context and disease model, with no universally standardized protocol; dosing is customized based on study goals, mouse strain, and model specifics.

Key details from published studies and supplier guidelines:

• Commonly reported doses range from 200 μg per mouse, administered intraperitoneally (i.p.), at regular intervals such as every 3 days for 4–6 doses.
• In a tumor model study, 200 μg per mouse was given i.p. every 3 days for four times starting after a treatment intervention (e.g., microwave ablation).
• Another study using lung metastasis models reported 200 μg per mouse i.p. every 3 days, repeated up to six times.
• Dosing can also be altered by tumor burden, mouse weight, or experimental timelines (e.g., after tumor engraftment, following immunotherapy initiation), and concurrent therapies may further influence the regimen.

Additional contextual considerations:

• Regimen must be tailored for the specific mouse model (e.g., tumor type, genetic background, immunological status).
• Suppliers (e.g., Bio X Cell, Thermo Fisher) and published literature should be consulted for guidance appropriate to the chosen model and objectives.
• Some studies do not specify the fractionation schedule, only the total dosing, so always refer to detailed methods sections for each experiment.

Summary Table: Typical Dosing Schedules for MIG-2F5-5 in Mouse Models

For highest accuracy, tailor dosing to the specific mouse model, experimental protocol, and consult supplier datasheets or primary publications for regimen justification.