Anti-Mouse IL-12 p75 [Clone R2-9A5] — Purified in vivo GOLD™ Functional Grade

The clone R2-9A5 is most commonly used in vivo in mice for neutralization of interleukin-12 (IL-12), specifically the p75 (p70) heterodimeric form of the cytokine. R2-9A5 is a rat IgG2b monoclonal antibody directed against mouse IL-12p75, which is composed of IL-12p40 and IL-12p35 subunits.

Key in vivo applications of R2-9A5 in mice include:

• Neutralization of IL-12 bioactivity: Administration of R2-9A5 effectively blocks IL-12 signaling, allowing researchers to examine the consequences of IL-12 pathway inhibition in settings such as infectious disease, autoimmunity, and tumor immunology.

• Immunomodulation studies: By inhibiting IL-12, R2-9A5 is used to investigate the role of this cytokine in T helper 1 (Th1) responses, cytotoxic T cell generation, and interferon-gamma production in vivo.

• Functional studies in immuno-oncology and inflammation: R2-9A5 helps dissect the contribution of IL-12 in models of cancer, infectious disease, and inflammatory disorders. For example, blocking IL-12 can reveal its impact on tumor immunity or the development of inflammatory lesions.

• Imaging applications: The antibody has been radiolabeled (e.g., with ^89Zr) for ImmunoPET imaging to non-invasively visualize sites of IL-12 expression and inflammation in live mice.

Typical experimental approaches include systemic injection of R2-9A5 to achieve functional blockade of IL-12, followed by assessments such as immune cell profiling, cytokine quantification, survival analysis, and imaging studies.

Summary Table: Common in vivo applications of clone R2-9A5 in mice

R2-9A5 is not typically used for cell depletion or as a detection reagent in flow cytometry, but is specifically chosen for its ability to neutralize IL-12 in functional studies.

The R2-9A5 antibody is primarily used to neutralize the biological effects of mouse IL-12, a critical cytokine involved in cell-mediated immune responses. While specific combinations of other antibodies or proteins used alongside R2-9A5 may not be explicitly detailed in the provided literature, several commonly used antibodies and proteins in immunological research could potentially be used in conjunction with it. These might include:

• Anti-mouse IL-1β antibodies: These are used to study inflammation and immune responses, similar to IL-12, and could be used in combination to explore broader immune modulation.
• Anti-mouse IFN-β antibodies: These are involved in interferon signaling and could complement IL-12 research by examining broader cytokine interactions.
• Anti-mouse IL-5 antibodies: These target eosinophil activation and growth, providing a different perspective on immune cell regulation.
• Anti-mouse IFNAR-1 antibodies: These are involved in interferon signaling and could be used with R2-9A5 to explore effects on immune response pathways.

These antibodies and proteins are commonly used in immunological studies and could be used in conjunction with R2-9A5 to explore various aspects of immune function. However, specific co-use cases might need to be identified through broader literature reviews.

Clone R2-9A5 is a monoclonal antibody widely cited in scientific literature for its ability to neutralize mouse interleukin-12 (IL-12), specifically the p75 (heterodimeric p40/p35) form, in both in vivo and in vitro experiments. The key findings from studies using R2-9A5 can be summarized as follows:

• Neutralization of IL-12: R2-9A5 binds and blocks the activity of IL-12, a cytokine that is critical for the differentiation of Th1 cells and the promotion of IFNγ and IL-2 production, thereby modulating cell-mediated immunity.

• Functional Impact in Disease Models:

  • In models of immunization and infection, administration of R2-9A5 allows researchers to dissect the role of IL-12 in Th polarization, immune activation, and disease pathogenesis.
  • For example, in studies of intradermal DNA vaccination, in vivo blockade with R2-9A5 showed that IL-12 is required for early Th1-associated immune responses. Cytokine profiles and transcription factor expression in T cells were altered following IL-12 inhibition, demonstrating the antibody’s utility in functional studies.
  • In skin inflammation models, manipulating IL-12 signaling (using neutralizing antibodies like R2-9A5) clarified IL-12's protective and regulatory role in limiting psoriasiform inflammation through effects in keratinocytes.

• Technical and Experimental Details:

  • R2-9A5 has been used in various experimental formats:
    • In vitro cocultures to assess T cell differentiation.
    • In vivo administration (typically 500 μg injected intraperitoneally) to block IL-12 function at specific time points in mouse models.
  • The antibody is a rat IgG2b, κ isotype, optimized for murine studies.

• Imaging and Quantitation:

  • R2-9A5 has been conjugated for use in immunoPET imaging of IL-12 distribution, enabling quantification and visualization of the cytokine in preclinical models.

In summary:
R2-9A5 is an authoritative IL-12 neutralizing tool in mouse immunology. Its citations reveal pivotal roles in:

• Defining the role of IL-12 in Th1 cell differentiation and immune polarization,
• Demonstrating the impact of IL-12 on disease outcomes and immune responses,
• Providing a reliable reagent for imaging and function-blocking studies in murine models.

While there is specific information available about the monoclonal antibody clone R2-9A5, which targets mouse IL-12 p75, general dosing regimens for antibodies in mouse models often vary based on the specific application and the model used. However, the R2-9A5 antibody is mentioned in a study where mice received 1 mg of this antibody via intraperitoneal (i.p.) injection once a week for three doses. Here's a broader overview of antibody dosing in mouse models:

• General Dosing Ranges: Antibody dosing in mouse models typically ranges from 100 μg to 500 μg per mouse, depending on the antibody and application.
• Common Routes: Intraperitoneal (i.p.) injection is a standard route for antibody administration in mouse models due to its convenience and effectiveness.
• Specific Examples:
  • RMP1-14 (Anti-PD-1): Often used at 200 μg per dose, with dosing schedules of every 3-4 days.
  • 10F.9G2 (Anti-PD-L1): Used at 100-250 μg per dose, with schedules of 2-3 times a week.

For clone R2-9A5, specific dosing might depend on the experimental design and the model's sensitivity to IL-12 neutralization. Therefore, while the dosing regimen for R2-9A5 might be similar to other antibodies in mouse models, the exact dosing could vary based on the specific research question and experimental conditions.