Anti-Mouse IFN-γ – Purified in vivo PLATINUM™ Functional Grade

The clone R4-6A2 is used in in vivo mouse studies primarily as a tool for blocking or neutralizing interferon-gamma (IFN-?), a cytokine critical for immune responses. This rat monoclonal antibody specifically binds to mouse IFN-?, which is a 20 kDa soluble cytokine produced mainly by activated lymphocytes, including T cells, B cells, NK cells, and ILCs. In vivo studies often utilize R4-6A2 to modulate IFN-?'s role in immune responses, such as in models of autoimmune diseases, infections, or cancer, where IFN-? plays a significant role in inflammation, cell differentiation, and immune regulation.

Applications in In Vivo Studies:

• Neutralization of IFN-? Activity: R4-6A2 can be administered to block the action of IFN-?, helping researchers understand its role in various biological processes and disease models.
• Immunological Studies: It is used to study the effects of IFN-? deficiency on immune responses, such as T cell activation and macrophage function.
• Cancer Research: R4-6A2 can be used to investigate the role of IFN-? in tumor growth and immune surveillance.

In vivo grade recombinant versions of R4-6A2 are available for these applications, ensuring compatibility and efficacy in mouse models.

The correct storage temperature for sterile packaged clone R4-6A2 (anti-mouse IFN-? antibody) is 2°C to 8°C when undiluted and for short-term use. For long-term storage, it may also be stored at -20°C or below to preserve stability.

• Multiple manufacturers specify 2°C to 8°C (standard refrigerator temperature) for the antibody solution in phosphate-buffered, preservative-free formulations.
• Alternative suppliers note that clone R4-6A2 can be stored at -20°C to -70°C for up to 12 months, especially for long-term storage, but must avoid repeated freeze-thaw cycles.
• When stored at 2°C to 8°C, expect typical antibody shelf life; if using frozen storage, thaw only once and aliquot as needed.

Key summary:

• Short-term: 2°C to 8°C (refrigerated).
• Long-term: -20°C or below (frozen, manual defrost freezer).
• Always consult the product datasheet/Certificate of Analysis for lot-specific instructions.

R4-6A2 is a widely used monoclonal antibody specific for mouse interferon-? (IFN-?), frequently employed as a capture or detection antibody in sandwich immunoassays such as ELISA and ELISpot. In the literature and assay protocols, R4-6A2 is commonly paired with other antibodies and proteins to enable sensitive and specific detection of IFN-?. Below are the most frequently cited partners:

Common Antibody Partners

• XMG1.2: This antibody is the most common partner for R4-6A2 in sandwich assays, serving interchangeably as either the capture or detection antibody depending on the protocol.
• AN18: For ELISpot and FluoroSpot, R4-6A2 is often paired with the AN18 antibody as the capture mAb, with R4-6A2-biotin used for detection.
• Recombinant Mouse IFN-?: Used as a standard curve protein in these assays to quantify IFN-? in samples.

Additional Notes

• Cross-reactivity: The R4-6A2/XMG1.2 pair does not cross-react with a wide panel of other cytokines, including mouse, human, and rat cytokines, ensuring specificity for mouse IFN-?.
• Application flexibility: R4-6A2 is also used for neutralization studies, but this application does not typically require a paired antibody.
• Other conjugates: While most protocols refer to biotinylated detection antibodies, other conjugates (e.g., fluorochromes for flow cytometry) are sometimes used with XMG1.2, but R4-6A2 is not typically described in flow cytometry applications.

Summary Table of Common Pairings

In summary, R4-6A2 is most commonly used with XMG1.2 for sandwich immunoassays, and with AN18 in ELISpot/FluoroSpot, while recombinant mouse IFN-? serves as the key standard protein for quantification. These pairings are standardized in most published protocols for mouse IFN-? detection.

Clone R4-6A2 is a rat monoclonal antibody widely used for detecting, quantifying, or neutralizing mouse interferon-gamma (IFN-?) in both in vitro and in vivo investigations. Key findings from scientific literature using this clone are summarized below:

• Neutralization of IFN-? in vivo: R4-6A2 is a standard tool for neutralizing IFN-? in mice to study the cytokine's biological functions. Neutralization experiments (typically via intraperitoneal injection) have uncovered critical roles of IFN-? in antiviral and antibacterial immunity, susceptibility to certain pathogens, and regulation of immune responses. For example, R4-6A2-mediated neutralization increased susceptibility to severe invasive Group A Streptococcus (GAS) infections, showing IFN-?’s protective effects against bacterial disease.

• Characterization in Immunoassays: R4-6A2 is a well-established reagent for ELISA, ELISPOT, flow cytometry, Western blot, and immunohistochemistry, being both a capture and detection antibody for mouse IFN-?. It is especially favored for quantifying IFN-? in murine samples.

• Functional Studies and Mechanistic Insights: By blocking IFN-?, R4-6A2 has been instrumental in revealing that IFN-? is involved in the regulation of cell-mediated immunity, activation of natural killer (NK) cells, and the orchestration of T-helper type 1 (Th1) responses. Its use highlighted mechanisms such as posttranscriptional control of cytokine expression and the importance of local (in situ) production of IFN-? for immune function.

• Role in B cell Biology: Studies using R4-6A2 indicate that IFN-? signaling, together with B-cell receptor (BCR) and TLR7 (a toll-like receptor), synergizes to induce high levels of T-bet expression in B cells. This finding links IFN-? to antibody class switching and antiviral responses.

• Extensive Citation and Standardization: The antibody’s specificity and performance are supported by numerous citations and longstanding use, including in defining studies (e.g., PMID:16386804). Its wide adoption has made it a gold standard for murine IFN-? detection and functional studies in immunology.

In summary, R4-6A2 has been fundamental in elucidating IFN-?'s immune functions in mice—especially its centrality to Th1 immunity, pathogen defense, and posttranscriptional regulatory mechanisms—and is an indispensable reagent for cytokine quantification and neutralization in experimental immunology.