Anti-Mouse IFNγ [Clone XMG1.2] — Purified in vivo PLATINUM™ Functional Grade

Clone XMG1.2 is most commonly used in vivo in mice to neutralize interferon gamma (IFNγ) function. This allows researchers to dissect the biological role of IFNγ in immune responses, inflammation, infection, autoimmunity, and tumor models by effectively blocking its activity during experimental studies.

Additional key in vivo applications include:

• Functional studies: Blocking IFNγ signaling to investigate its contribution to pathogenesis, host defense, and immune regulation. This is used in models of infection (viral, bacterial, parasitic), tumor immunity, autoimmunity, and inflammation.
• Cytokine profiling and immune cell characterization: While more common ex vivo or in vitro, the antibody may be administered to mice to facilitate the downstream detection of IFNγ-producing cells or analysis of systemic cytokine production, typically using flow cytometry or ELISA post-sample collection.
• Dissecting mechanisms of therapies or disease processes: Neutralization studies can help determine whether therapeutic efficacy or pathology in a given mouse model is IFNγ dependent.

Compared to in vitro uses (such as ELISA, flow cytometry, and ELISPOT), in vivo use of XMG1.2 is primarily for neutralizing IFNγ to elucidate its physiological roles. For in vivo studies, endotoxin-free formulations are recommended to minimize adverse immune activation unrelated to IFNγ blockade.

In summary, the principal in vivo application of clone XMG1.2 in mice is neutralization of IFNγ bioactivity to study its role in immune processes and disease models.

Commonly used antibodies or proteins in the literature used with XMG1.2 (anti-mouse IFN-γ) typically target other cytokines or immune cell markers for multiparameter immune profiling, or they are part of ELISA/ELISPOT assay pairs for IFN-γ detection.

Key antibodies and proteins commonly used with XMG1.2 include:

• R4-6A2 Antibody: This is the most frequently paired antibody with XMG1.2 for ELISA and ELISPOT. In sandwich assays, XMG1.2 is often used as the capture antibody while R4-6A2 is used as the detection (biotinylated) antibody, or vice versa.
  Example (ELISA/ELISPOT pairing):
  | Capture Antibody | Detection Antibody ||------------------|---------------------------|| XMG1.2 | Biotinylated R4-6A2 || R4-6A2 | Biotinylated XMG1.2 |

• Recombinant Mouse IFN-γ Protein: Used as a standard for quantification in ELISA or ELISPOT assays alongside XMG1.2.

• Isotype Controls: Purified Rat IgG1, κ isotype controls are routinely used for background correction and ensuring specificity when XMG1.2 (rat IgG1, κ) is used in flow cytometry or in vivo experiments.

• Other Cytokine Antibodies: In multiparametric flow cytometry or immunoassays, XMG1.2 is often used in combination with antibodies against:

  • IL-2 (anti-IL-2; JES6-5H4 clone)
  • IL-4 (anti-IL-4; 11B11 clone)
  • IL-17A (anti-IL-17A; TC11-18H10.1 clone)
  • TNF-α (anti-TNF; MP6-XT22 clone)
  • These combinations allow identification of T helper cell subsets and polyfunctional T cell responses, especially in studies of infection, autoimmunity, and cancer.

• Cell Surface Markers: For immune cell phenotyping, XMG1.2 is frequently used in combination with antibodies targeting markers such as:

  • CD3, CD4, CD8 (T cell markers)
  • CD45, CD11b (leukocyte/myeloid markers)
  • These are used in multicolor flow cytometry panels.

• Fluorochrome Conjugates: XMG1.2 is available conjugated to many fluorochromes (e.g., FITC, PE, APC, PerCP/Cy5.5, Brilliant Violet 480), allowing combination with other fluorescent antibodies for multicolor flow cytometry panels.

Summary Table: Commonly Paired Reagents with XMG1.2

These combinations are widely supported by both product application guides and peer-reviewed experimental protocols. For specific panel designs or protocols, refer to assay kit documentation or recent immunology publications using XMG1.2.

Clone XMG1.2 is a rat monoclonal antibody widely used in scientific research for the detection and neutralization of mouse interferon-gamma (IFN-γ), a key cytokine in immune regulation. The major findings from XMG1.2 citations in the literature are:

• Biological Understanding of IFN-γ: Studies using clone XMG1.2 have established that IFN-γ is a pleiotropic cytokine, critically involved in immune regulation, upregulation of major histocompatibility complex (MHC) molecules, macrophage activation, and modulation of lymphocyte functions.
• Antibody Properties and Applications: XMG1.2 specifically binds mouse IFN-γ and is highly cited for:
  • Intracellular cytokine staining in flow cytometric analysis to detect activated T cells.
  • ELISA assays: Forms a detection pair with clones such as R4-6A2 to quantify IFN-γ.
  • Neutralization experiments: Used to block IFN-γ function in vivo and in vitro, helping characterize its role in immune responses.
  • Immunofluorescence and immunohistochemistry for tissue analysis.
• Optimization of Antibody Fragmentation: Studies focusing on enzymatic fragmentation of XMG1.2 revealed:
  • F(ab')₂ fragments generated from XMG1.2 can exhibit higher antigen-binding affinity compared to the intact antibody, possibly due to increased hinge flexibility after Fc removal.
  • Optimal fragmentation conditions (pH, enzyme concentration) boost yield and affinity, with low pH causing undesirable aggregation.
• Functional Role of IFN-γ in Disease Models: Using XMG1.2, research has shown that:
  • IFN-γ limits the expansion of type 2 lymphocytes (such as ILC2 and Th2 cells) in allergic inflammation.
  • IFN-γ contributes to antiviral, antitumor responses, and shapes tissue immune niches by constraining certain immune cell infiltrations.
• In Vivo and In Vitro Use: XMG1.2 is frequently employed in mouse model experiments to dissect IFN-γ’s role in infection, cancer, and autoimmunity through both detection and neutralization approaches.

Summary Table of Key Findings with Clone XMG1.2

In summary, citations of clone XMG1.2 have been pivotal in defining the biological roles of IFN-γ, especially in immune modulation and disease, as well as providing technical advancements in antibody-based detection and functional analysis methods.

Dosing Regimens of Clone XMG1.2 in Mouse Models

Overview
Clone XMG1.2 is a widely used neutralizing monoclonal antibody targeting mouse interferon-gamma (IFNγ). Its dosing regimens in vivo vary significantly depending on the specific mouse model, disease context, and desired biological effect (e.g., neutralization, depletion, or functional blockade).

Model-Specific Dosing

Key Observations

• Dose Dependency: Higher doses (30–100 mg/kg) are generally required for effective neutralization of IFNγ in inflammatory or hypercytokinemia models, such as hemophagocytic lymphohistiocytosis (HLH) or TLR9-driven cytokine storms. Lower doses may be insufficient to significantly alter disease parameters in certain contexts.
• Route of Administration: Intraperitoneal injection is common, but intravenous administration has also been used in specific studies.
• Frequency: Dosing frequency varies from single administration to repeated doses over days or weeks, tailored to the kinetics of the disease or experimental endpoint.
• Model-Specific Variability: The same dose may have different effects in different models. For example, 8 mg/kg was ineffective in a cytomegalovirus-driven HLH model but higher doses were effective in LCMV-driven HLH.
• Use Outside Hyperinflammation: In models where the goal is modest immunomodulation (e.g., vaccine studies), much lower doses (e.g., 200 μg/mouse) have been used.

Summary Table: Representative Dosing Examples

Conclusion

The dosing regimen of clone XMG1.2 in mouse models is highly context-dependent. Higher doses (30–100 mg/kg) are typically required for robust IFNγ neutralization in hyperinflammatory disease models, while lower doses (e.g., 200 μg/mouse) may suffice for modulating immune responses in less severe settings. Researchers should carefully consider the disease context, desired biological effect, and model-specific literature when designing XMG1.2 dosing regimens.