Anti-Human IL-4 – Purified in vivo GOLD™ Functional Grade

Clone MP4-25D2 is most commonly used in vivo to neutralize human IL-4 in mouse models for studying immune modulation and cytokine function, particularly in xenograft or humanized mouse systems.

This antibody is specific for human IL-4, a cytokine involved in Th2 cell differentiation and B-cell responses. Its principal in vivo applications include:

• Neutralization of human IL-4: MP4-25D2 is used to block human IL-4 activity after engraftment of human cells into mice or in humanized mouse models, enabling researchers to assess the effects of IL-4 inhibition on immune responses, allergy, autoimmunity, or tumor growth. This approach helps to dissect the role of IL-4 in disease processes involving human cytokines.
• Functional studies: By injecting MP4-25D2 in vivo, investigators can modulate Th2-type immune responses driven by human IL-4 and study downstream effects on cell populations or disease symptoms.

Note: MP4-25D2 does not cross-react with mouse IL-4—it is specific for human IL-4, so in vivo applications are limited to models expressing human IL-4, such as human cell xenografts or transgenic mice. For mouse IL-4 studies, other clones (e.g., 11B11) are used.

Additional relevant methods for MP4-25D2 in research include in vitro neutralization, flow cytometry, ELISA, immunohistochemistry, and immunofluorescence, but direct in vivo use in mice centers on neutralizing human IL-4 in settings where human cytokine function is analyzed within a living mouse organism.

Alongside MP4-25D2, which targets human IL-4, other commonly used antibodies or proteins in the literature include those directed against T cell, B cell, and cytokine markers, as well as other IL-4 antibodies used in combination assays.

Key proteins and antibodies frequently used with or in similar experiments as MP4-25D2 include:

• CD3 antibody: Used as a T cell marker, frequently for co-staining in flow cytometry to identify T cell subsets producing IL-4.
• CD4 antibody: Utilized for identifying helper T cells (especially Th2) that produce IL-4.
• CD45RA: Surface marker for distinguishing naïve vs. memory T cell subsets in studies tracking IL-4 production.
• Other anti-IL-4 clones: Most notably, the 8D4-8 clone is widely used for IL-4 detection (often paired as the capture or detection antibody in ELISA/ELISpot with MP4-25D2 or used in parallel flow cytometry panels). Additional anti-IL-4 clones include 4D9 and REA895.
• Isotype controls: Such as rat IgG1, to validate staining specificity in flow cytometry panels.
• Cytokines and stimulation cocktails: Including recombinant human IL-2 and IL-4 (for T cell stimulation), PMA and ionomycin (for robust cytokine induction), and GolgiStop™ to block cytokine secretion.
• Other immunophenotyping antibodies (for multi-parametric flow cytometry): For example, anti-CD19 (B cell marker), anti-CD8 (cytotoxic T cells), anti-IFNγ, or anti-IL-2, depending on the experimental focus.

In addition to these, MP4-25D2 is often paired with biotinylated or fluorochrome-conjugated forms of itself or other IL-4 antibodies, particularly in ELISA and intracellular staining protocols.

In summary, MP4-25D2 is frequently used in concert with CD3, CD4, CD45RA, isotype controls, cytokine stimulation agents, and other anti-IL-4 clones (especially 8D4-8) for multi-parameter immunoassays targeting human T cell cytokine responses.

Clone MP4-25D2 is a rat monoclonal antibody widely used in scientific literature to specifically detect and neutralize human interleukin-4 (IL-4) in both basic and translational immunological research. The key findings from citations involving this clone are:

• Specificity for IL-4: MP4-25D2 has high specificity for human IL-4, a crucial cytokine in Th2 immune responses, B cell proliferation, and differentiation.

• Applications in Intracellular Cytokine Staining: MP4-25D2 is prominently used for intracellular staining of IL-4 in human peripheral blood mononuclear cells (PBMCs), especially after stimulation with agents such as PMA and ionomycin, followed by fixation, permeabilization, and staining with the fluorochrome-conjugated antibody.

• Validation of Specificity: The specificity is frequently demonstrated by blocking experiments where excess unlabeled MP4-25D2 antibody inhibits binding of its fluorochrome-conjugated counterpart, confirming signal accuracy in flow cytometry and excluding nonspecific interactions.

• Neutralization Capability: MP4-25D2 can neutralize the biological activity of both recombinant and natural IL-4, making it useful for functional studies investigating IL-4's role in cell signaling and immune responses. For example, it has been used to completely neutralize IL-4 in in vitro co-culture experiments with CAR-T cells, highlighting its effectiveness in modulating cytokine-driven processes.

• Negligible Non-specific Binding: Compared to some other IL-4 antibody clones, MP4-25D2 shows negligible off-target binding, such as non-cross-reactivity with other cellular components or live/dead discrimination reagents in flow cytometry.

• Broad Citation in Methods: MP4-25D2 is a gold-standard reagent for studying Th2-type immune responses and for measuring IL-4 expression in a range of experimental contexts, providing well-documented reliability and reproducibility in literature.

• Commercial Availability: The antibody is available from multiple suppliers in different conjugated formats (e.g., PE, purified), supporting diverse research protocols and reproducibility standards.

In summary, MP4-25D2 is a widely validated, highly specific, and functionally neutralizing antibody for human IL-4, with key applications in intracellular cytokine detection, functional neutralization assays, and flow cytometric quantification of Th2 immune responses. It is frequently cited for its reliability and minimal cross-reactivity in both basic and translational immunology experiments.

The dosing regimens of clone MP4-25D2—a monoclonal antibody specific for human IL-4—are not standardized across different mouse models, primarily because its primary use is for detection (e.g., flow cytometry, ELISA, immunostaining) rather than therapy, and the details reported in the literature pertain almost exclusively to in vitro assays and ex vivo cell analysis rather than in vivo dosing in mice.

Key details from available sources:

• Application usage: MP4-25D2 is routinely used for intracellular staining, often at doses like 0.05 µg per staining reaction (for ~1 million cells), and as a blocking antibody in flow cytometry protocols where unlabeled antibody can be used at higher concentrations, such as 2.5 µg per reaction for specificity controls.
• Species reactivity: The MP4-25D2 clone is rat anti-human IL-4, with reported cross-reactivity with rhesus monkey IL-4, but is not reported to bind mouse IL-4.
• In vivo use: There is no evidence in the search results that MP4-25D2 is used for in vivo dosing in mouse models (i.e., as a treatment or for functional neutralization in mice). Its applications are consistently reported in the context of detection of human IL-4 in cell cultures or in humanized mouse models where human cells have been engrafted.

In humanized mouse models:

• MP4-25D2 may be used for ex vivo analysis of engrafted human cells (such as post-transfer to NOD/scid mice, as in Treg cell functional studies), again at reagent concentrations defined by staining protocols, not therapeutic dosing.

Summary Table: Dosing Regimens by Use-case

Important context:

• MP4-25D2 is not used as a therapeutic in mouse models, and due to its target specificity (human IL-4), it’s not applicable for blockade of endogenous mouse IL-4 pathways.
• Detection protocols are similar regardless of mouse strain, as they depend on the presence of human IL-4-expressing cells.

If you require information on dosing of anti-mouse IL-4 antibodies or therapeutic regimens instead, those would involve different clones and regimens that are species-matched and have published dosing protocols.