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

Clone XMG1.2 is a monoclonal antibody that specifically binds to mouse interferon gamma (IFN?) and is widely used in in vivo mouse studies to neutralize or block the activity of endogenous IFN?, thereby modulating immune responses.

In context:

• Neutralization/Inhibition: The primary in vivo application of XMG1.2 is to functionally neutralize IFN? in murine models. This allows researchers to dissect the role of IFN? in various contexts such as infection, inflammation, autoimmune diseases, and vaccine responses.
• Preclinical Disease Models: By blocking IFN? signaling in live mice, the antibody is used to study:
  • The effects of IFN? on cytotoxic T-cell (CTL) responses, as shown in vaccine immunogenicity studies where anti-IFN? XMG1.2 enhanced CTL responses to candidate vaccines.
  • Mechanistic roles of IFN? in infectious disease, allergy, cancer, and transplantation models.
• Species-Matched Variants: Recombinant chimeric versions (e.g., XMG1.2-CP058, rat variable domains with mouse constant regions) are available for use in mice to minimize immunogenicity and anti-drug antibody formation, improving reliability and reproducibility compared to traditional rat anti-mouse antibodies.
• Functional Grade Purification: Ultra-low endotoxin preparations (e.g., "Ultra-LEAF™") are recommended for in vivo work, further reducing confounding inflammatory effects from contaminants.

Additional technical uses:

• While XMG1.2 can also be employed for detection of IFN? by flow cytometry or ELISA, its defining use in vivo is functional blockade of IFN? signaling in mouse models to examine how immune or pathological processes depend on this cytokine.

Summary of key applications:

• Neutralizing mouse IFN? in living mice to study immune function.
• Modulating immune responses during disease or therapy.
• Reducing formation of anti-drug antibodies by using species-matched or recombinant forms for chronic studies.

XMG1.2 is specifically recommended over other clones when high specificity and effective neutralization of mouse IFN? are needed for mechanistic studies in vivo.

Sterile, packaged clone XMG1.2 monoclonal antibody should be stored at 2°C to 8°C and must not be frozen.

Multiple sources specify that the antibody solution, including clone XMG1.2, ought to be kept undiluted between 2°C and 8°C and protected from light exposure. Freezing is explicitly discouraged, as this may compromise the antibody's activity and stability. If lyophilized (freeze-dried), storage at -20°C to -70°C may be appropriate, but once reconstituted, the storage recommendation reverts to 2–8°C for short to medium-term use.

• For immediate use (weeks to a few months): 2°C–8°C, protected from light, do not freeze.
• For lyophilized antibodies: Store desiccated at -20°C to -70°C until reconstituted; then follow the above short-term protocol.

Always confirm with the specific product datasheet or manufacturer for clone XMG1.2, as conjugated forms or formulations (e.g., with preservatives/carriers) may have additional requirements.

Commonly used antibodies or proteins paired with XMG1.2 (anti-mouse IFN-?) in the literature include:

• Anti-IL-4 (e.g., clone 11B11): Frequently used in multiparameter flow cytometry or ELISA panels to profile Th1/Th2 immune responses alongside IFN-? detection.
• Anti-IL-2: Often co-stained to characterize functional subsets of T cells (especially helper T cell responses).
• Anti-TNF-?: Used to monitor polyfunctional T cells and the broader inflammatory cytokine profile.
• Surface markers: Commonly include
  • CD4 (e.g., GK1.5) and CD8 (e.g., 53-6.7) for identifying T cell subsets producing IFN-?.
  • CD3 (e.g., 145-2C11) to confirm T cell lineage.
  • CD19 for B cells when IFN-? production from other lymphoid sources is studied.
• Viability dyes: Included to exclude dead cells from analysis in flow cytometry experiments.
• Capture/detection pairs: In ELISA, XMG1.2 is regularly paired with R4-6A2 as the capture antibody, and recombinant mouse IFN-? as a standard protein.

These antibodies and proteins are selected based on the assay and research objective:

• In ELISA, XMG1.2 is used as a detection antibody, often in combination with R4-6A2 as a capture antibody.
• For intracellular cytokine staining (ICS) panels in flow cytometry, XMG1.2 may be combined with anti-IL-4, anti-IL-2, anti-TNF-?, and appropriate surface markers to profile T cell cytokine production and phenotype.

In summary, when studying mouse IFN-? with XMG1.2, researchers frequently use antibodies against other cytokines (IL-2, IL-4, TNF-?) and surface lineage markers (CD4, CD8, CD3), especially in flow cytometry and ELISA protocols, to provide a comprehensive immune profile.

Clone XMG1.2 is a rat monoclonal antibody widely used in scientific literature for detecting and neutralizing mouse interferon-gamma (IFN-?), with key findings across immunological research and disease models.

Key findings from XMG1.2 citations include:

• Detection and Quantification of IFN-?:
  XMG1.2 is routinely used for detection of intracellular and secreted IFN-? in mouse tissues and cells via flow cytometry, ELISA, and ELISPOT assays. This enables quantitative analysis of immune responses, particularly those involving activated T cells and other lymphocytes.

• Neutralization of IFN-? Biological Activity:
  The antibody is used to neutralize IFN-? both in vitro and in vivo, allowing investigators to assess the specific role of IFN-? in a variety of contexts such as infection, autoimmunity, and neuroinflammation. Neutralization studies using XMG1.2 have shown its effectiveness in suppressing IFN-?-mediated effects, providing evidence for the cytokine’s pathological and regulatory functions.

• Role in Disease Models:
  In a recent neuroinflammatory study, administration of XMG1.2 in a mouse model of multiple system atrophy (MSA) attenuated pro-inflammatory monocyte infiltration, decreased MHCII expression on microglia, and reduced CD4+ and CD8+ T cell entry into the brain. This established IFN-? as a mediator of ?-synuclein-induced neuroinflammation and demyelination, and suggested IFN-? as a possible disease-modifying therapeutic target.

• Validation and Specificity:
  XMG1.2’s specificity is routinely validated in experimental settings—for example, staining specificity can be blocked by the unlabeled antibody, demonstrating its selectivity for IFN-? in mouse samples.

• Breadth of Applications:
  Beyond flow cytometry and neutralization, the antibody is cited for use in western blotting, immunohistochemistry, and immunocytochemistry, covering a wide array of investigative techniques for studying IFN-? biology. It is among the most frequently used clones for IFN-? in mouse research, with hundreds of citations.

Additional relevant insights:

• IFN-? Immunobiology:
  IFN-? is a central cytokine in immune regulation, anti-viral defense, and inflammation—XMG1.2 thus serves as a foundational reagent for elucidating immune mechanisms in mouse models.

• Translational Potential:
  Studies using XMG1.2 have helped identify therapeutic strategies targeting IFN-?, particularly for neurodegenerative and autoimmune diseases where IFN-? plays a pathogenic role.

In summary, clone XMG1.2 is a critical antibody for dissecting IFN-?’s role in mouse immunology, underpinning findings on immune response characterization, disease pathology, and therapeutic intervention strategies.