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

The MIG-2F5-5 clone is an anti-mouse CXCL9 monoclonal antibody primarily used for in vivo studies in mice to neutralize the chemokine CXCL9 (also known as monokine induced by gamma interferon, MIG). Its main application is to functionally block CXCL9 in order to study immune cell migration, inflammation, and related processes in living animals.

Key details relevant to in vivo mouse studies:

• Target & Specificity: MIG-2F5-5 is highly specific for murine CXCL9, a chemokine secreted by cells such as macrophages, monocytes, endothelial cells, fibroblasts, cancer cells, and especially dendritic cells in response to IFN-?.
• Isotype & Source: Armenian hamster IgG, produced from immunization with mouse plasmacytoid dendritic cells.
• Formulation: Provided as a sterile, low- or ultra-low-endotoxin IgG solution (typically <0.5 or <1.0 EU/mg), BSA and azide free, making it suitable for in vivo administration in mice.
• Purity: High purity (>95%–98% by SDS-PAGE/HPLC) to minimize potential off-target or immunogenic effects.
• Main Application in vivo: Used to neutralize endogenous CXCL9 in living mice, enabling researchers to dissect the role of this chemokine in immune cell trafficking, tumor immunity, autoimmune diseases, and tissue inflammation. This is accomplished by injecting the antibody into mice to block CXCL9 activity and then monitoring biological outcomes such as immune cell infiltration, disease progression, and related immune responses.
• Controls: Experiments typically include isotype control antibodies to confirm specificity of the observed effects.
• Pathogen Testing: Ultra-low endotoxin preparations are tested to be pathogen-free for common rodent pathogens to ensure animal safety.

In summary, clone MIG-2F5-5 is a high-purity, low-endotoxin antibody designed and validated for functional in vivo neutralization of murine CXCL9, allowing researchers to explore the specific roles of CXCL9 in experimental mouse models.

Storage Temperature for Sterile Packaged Clone MIG-2F5-5

• Short-Term Storage (Up to 1 Month): Functional grade, sterile, preclinical antibodies of clone MIG-2F5-5 can be stored as received at 2–8°C (refrigerated, not frozen) if they remain sterile and unopened.
• Long-Term Storage: For storage beyond one month, aseptically aliquot the antibody in working volumes (without dilution) and store at ?70°C. Avoid repeated freeze-thaw cycles to maintain antibody integrity.
• General Storage Guidance: Most suppliers (including BioLegend and Bio X Cell) of the MIG-2F5-5 clone recommend storing the antibody solution undiluted at 2–8°C and not freezing the stock solution unless specifically indicated for longer-term storage. Freezing is generally not recommended for the stock unless you need to extend storage beyond a few weeks.
• Conjugated Antibodies (e.g., PE-conjugated): Special conjugated forms may have additional requirements, such as protection from light, but the temperature recommendation (2–8°C) remains consistent.

Summary Table

Always check the specific datasheet or product insert for your lot, as recommendations may change.

Commonly Used Antibodies and Proteins Paired with MIG-2F5-5

The MIG-2F5-5 antibody specifically targets murine CXCL9 (also known as MIG, Monokine Induced by Gamma interferon). In the literature and research contexts, MIG-2F5-5 is often used to study the CXCL9/CXCR3 axis and its role in immune cell migration, differentiation, and activation—particularly in the context of tumor immunology, inflammation, and infection.

Key Partners in the CXCL9/CXCR3 Axis

• CXCL9, CXCL10, and CXCL11: These three chemokines, along with their common receptor CXCR3, are frequently studied together. They are known to regulate immune cell migration (especially Th1 cells), differentiation, and activation, and have been implicated in both tumor suppression and, in certain contexts, tumor promotion.
• CXCR3: The receptor for CXCL9, CXCL10, and CXCL11 is often examined alongside MIG-2F5-5 to understand chemokine signaling and immune cell recruitment.
• Interferon-gamma (IFN-?): Since CXCL9 expression is strongly dependent on IFN-?, this cytokine is a common co-factor in studies investigating CXCL9 regulation and function.
• Anti-TIM-3 and Anti-Galectin-9 Antibodies: In some experimental settings, MIG-2F5-5 has been used in conjunction with antibodies against immune checkpoint molecules like TIM-3 and Galectin-9. For example, neutralization of Galectin-9 or TIM-3 can enhance CXCL9 expression, which can be detected using MIG-2F5-5.

Cell Types and Markers

• Macrophages, Monocytes, Endothelial Cells, Fibroblasts, and Cancer Cells: These are the primary producers of CXCL9 in response to IFN-?, and thus are often the focus of studies using MIG-2F5-5.
• Intratumoral Dendritic Cells: CXCL9 is also detectable in specific subsets of dendritic cells within tumors, making dendritic cell markers (e.g., CD103+, CD8?+) relevant in these studies.
• CD103+ Conventional Dendritic Cells (cDCs): These have been shown to express CXCL9 in certain tumor models, especially when stimulated in vivo.

Experimental Contexts

• Flow Cytometry (FC) and Immunofluorescence (IF): MIG-2F5-5 is used for detecting CXCL9 expression at the protein level in these assays.
• Neutralization Studies: The antibody can be used to neutralize CXCL9 activity in vivo or in vitro to study its functional role.
• Combination with Other Cytokine/Chemokine Antibodies: In multiplex assays, MIG-2F5-5 may be paired with antibodies against other cytokines or chemokines (e.g., CXCL10, CXCL11) to profile immune responses.

Summary Table

Conclusion

MIG-2F5-5 is most commonly used alongside antibodies and proteins involved in the CXCL9/CXCR3 axis (CXCL10, CXCL11, CXCR3), IFN-?, and immune checkpoint molecules like TIM-3 and Galectin-9. It is also frequently paired with cell-type-specific markers to identify the cellular sources of CXCL9 in various experimental models. These combinations are central to investigating the role of CXCL9 in immunity, cancer, and inflammation.

Clone MIG-2F5-5 is a well-characterized monoclonal antibody targeting mouse CXCL9 (monokine induced by gamma interferon, MIG) that has been extensively used in scientific research to understand immune cell dynamics and therapeutic interventions. The key findings from scientific literature utilizing this clone reveal several important discoveries about CXCL9's role in immune responses and disease processes.

Anti-Rejection Therapy in Transplantation

One of the most significant findings involves the use of MIG-2F5.5 in combination therapy for preventing organ transplant rejection. Research has demonstrated that the combination of CXCR3-173 monoclonal antibody with MIG-2F5.5 monoclonal antibody could limit acute rejection significantly more than single neutralizing antibody approaches. This finding suggests that targeting both the CXCL9 chemokine and its receptor CXCR3 provides synergistic protection against transplant rejection, highlighting the importance of the CXCL9/CXCR3 axis in allograft immunity.

Cancer Immunotherapy and Metastasis Control

Perhaps the most compelling findings come from cancer research, where MIG-2F5.5 has been instrumental in demonstrating CXCL9's critical role in anti-tumor immunity. Studies using this antibody have shown that IFN?2-induced T cell recruitment in tumors depends specifically on CXCL9. When researchers used MIG-2F5.5 to neutralize CXCL9 in tumor-bearing mice treated with lipid nanoparticle-delivered IFN?2, they observed significant reductions in the therapy's effectiveness.

Metastasis Suppression Mechanisms: The research revealed that LNP-IFN?2 therapy suppresses tumor lung metastasis through upregulating CXCL9, and this effect is abolished when CXCL9 is neutralized with MIG-2F5.5. In both 4T1 mammary carcinoma and B16-F10 melanoma models, treatment with the anti-CXCL9 antibody significantly reduced the anti-metastatic effects of IFN?2 therapy, demonstrating CXCL9's essential role in recruiting tumor-infiltrating T cells.

T Cell Infiltration: Flow cytometry analysis using MIG-2F5.5-treated mice showed that neutralizing CXCL9 substantially reduced both CD4+ and CD8+ T cell infiltration into lung metastases. This finding establishes CXCL9 as a key mediator of T cell recruitment to tumor sites and explains why IFN?2 therapy loses its effectiveness when this chemokine is blocked.

Immune Cell Characterization

Single-cell RNA sequencing studies have utilized MIG-2F5.5 to characterize different T cell populations and their functional states. The antibody has been valuable in studying alloreactive T cell subsets, including highly proliferative (Mki67^hi^) cytotoxic T lymphocytes and chemokine-producing (Ccl5^hi^) CTLs in transplant rejection models. These studies have provided insights into the heterogeneity of immune responses and the specific roles of different T cell populations in allograft rejection.

Clinical Implications and Therapeutic Potential

The research findings using MIG-2F5.5 have revealed that in human cancer patients, IFN-I pathway activation correlates with CXCL9 expression and T cell expansion following PD-(L)1 immune checkpoint inhibitor immunotherapy. This correlation suggests that CXCL9 levels could serve as a biomarker for immunotherapy response and that strategies targeting the CXCL9 pathway could enhance current cancer treatments.

The studies demonstrate that LNP-encapsulated IFN?2 represents a potentially safe and effective third-generation IFN?2 agent for human cancer immunotherapy, particularly for treating patients with lung metastasis. The mechanism involves restoring IFN?2 expression to activate CXCL9 expression, which enhances T cell tumor recruitment to suppress metastasis.

These findings collectively establish MIG-2F5.5 as not just a research tool but as a key reagent that has helped elucidate CXCL9's central role in immune surveillance, transplant rejection, and cancer immunotherapy. The research demonstrates that CXCL9 serves as a critical bridge between innate immune activation and adaptive T cell responses, making it an important target for therapeutic intervention.