Anti-mouse/rat MHC Class II (I-Ek/RT1-D) (Clone 14-4-4S (HB-32)) – Purified in vivo GOLD™ Functional Grade

Clone 14-4-4S is a versatile monoclonal antibody widely used in mouse research for manipulating and studying MHC Class II molecules. This antibody specifically targets the I-Eᵏ MHC Class II alloantigen expressed in certain mouse strains (H-2ᵈ, H-2ᵖ, and H-2ʳ haplotypes) and also cross-reacts with rat RT1-D.

The primary in vivo applications of clone 14-4-4S include blocking antigen presentation and immune cell depletion studies. The antibody has been reported to block antigen presentation in functional assays, making it valuable for investigating the role of MHC Class II molecules in immune responses. Both whole antibody and F(ab')₂ fragments effectively block RT1-D-restricted antigen presentation as well as activation of I-E'-restricted T cell hybridomas.

For cell depletion studies, 14-4-4S enables researchers to selectively target specific immune cell populations. The antibody can be used for complement-mediated cell depletion, allowing researchers to eliminate B cells and other MHC Class II-expressing cells in vivo. Since I-Eᵏ and RT1-D are expressed primarily on B lymphocytes, antigen-presenting cells, macrophages, dendritic cells, and a subset of T cells, this antibody provides a tool for studying the functional roles of these populations.

The 14-4-4S antibody has also been reported to induce differentiation of mouse cells expressing I-Eᵏ, adding another dimension to its utility in immunological research. The antibody is typically formulated in low endotoxin/azide-free preparations specifically designed for in vivo use, ensuring compatibility with live animal studies.

Commonly Used Antibodies and Proteins with 14-4-4S in Literature

14-4-4S is a widely recognized monoclonal antibody primarily used to detect mouse and rat MHC class II (I-Ek) antigens expressed on professional antigen-presenting cells such as B cells, monocytes, macrophages, and dendritic cells. In immunological research, especially flow cytometry, co-staining and multiplex assays often involve other antibodies and signaling proteins. However, the literature primarily describes 14-4-4S alone or in combination with general cellular markers—not with a specific, recurrent set of antibodies or proteins—in most references. Below is a breakdown of the evidence and context.

Typical Experimental Context for 14-4-4S

• Cellular localization: 14-4-4S is used to identify MHC class II-positive cells, so it is often paired with antibodies against surface markers for the same cell types (e.g., B cells—CD19, CD45R/B220; monocytes/macrophages—CD11b, F4/80; dendritic cells—CD11c).
• Experimental applications: The antibody is validated for flow cytometry, immunoprecipitation, immunohistochemistry, and complement-dependent cytotoxicity.
• Functional assays: 14-4-4S can block antigen presentation in vitro, so it may be used in studies of antigen processing and presentation.

Commonly Co-Used Antibodies and Proteins

While no single, high-frequency molecular partner for 14-4-4S is identified in the provided literature, in the broader context of MHC class II research, the following are commonly co-detected or functionally relevant:

These pairings are inferred from standard immunological practice and the cell types known to express MHC class II, rather than from explicit citations in the provided references.

14-4-4S-Specific Literature Evidence

• Idiotype studies: There is one citation that used a heterologous anti-idiotypic reagent with 14-4-4S, but this is not a typical experimental antibody pair; rather, it is for characterizing the idiotype of 14-4-4S itself.
• Cross-reactivity: 14-4-4S cross-reacts with rat MHC class II (RT1D), so in rat studies, analogous rat cell markers may be used.
• No signaling partners: No literature in the provided search results mentions direct interaction partners (e.g., adaptors, signaling proteins) for 14-4-4S itself; this contrasts with, for example, antibodies against signaling molecules (like phospho-STATs, ZAP-70, etc.) in other immunological assays.

Summary

14-4-4S is most often used as a stand-alone reagent to detect MHC class II (I-Ek) on murine or rat antigen-presenting cells. In multiplex assays, it is routinely paired with markers for B cells (CD19, B220), myeloid cells (CD11b, F4/80), and dendritic cells (CD11c) to characterize immune cell subsets and their activation states, but no unique, high-frequency molecular partners for 14-4-4S are specified in the literature. For functional blocking studies, 14-4-4S is noted for inhibiting antigen presentation, but this does not imply interaction with other proteins or antibodies beyond its target antigen.

If you have a specific experimental context (e.g., autoimmune models, vaccination, or infection), additional customized antibody panels may be used, but these are not documented in the general product literature or review citations provided. For detailed protocol examples, consulting cell-type-specific marker panels in immunology methods resources would be advisable.

Clone 14-4-4S is a monoclonal antibody widely cited in scientific literature for its specificity to the mouse MHC class II I-E subregion-encoded glycoprotein, with key findings centered on its use in immunology research:

• Specificity and Shared Idiotype: 14-4-4S is highly specific for the I-E antigen, particularly the Ia.7 determinant, and recognizes mature MHC class II heterodimers (I-E) on the surface of antigen-presenting cells in mice with H-2^k, H-2^d, H-2^p, and H-2^r haplotypes. Some strains, such as H-2^b, H-2^f, and H-2^s, do not display I-E antigens and are not recognized by this clone. Importantly, 14-4-4S expresses a shared idiotype, making it a model for studying idiotype networks and immune regulation.

• Functional Applications: The antibody is commonly used for:

  • Flow cytometry to identify and quantify I-E–positive cells.
  • Immunoprecipitation and immunohistochemistry of class II MHC molecules.
  • Blocking antigen presentation in vitro, thus enabling the study of antigen processing and T cell activation.
  • Cytotoxicity assays owing to its proven activity in complement-dependent lysis of I-E–positive cells.

• Biochemical Insights: The antibody exclusively detects mature I-E molecules on the cell surface, not those associated with invariant chains or unassembled forms. This has helped define the cellular trafficking and maturation of MHC class II molecules, showing that after B cell activation, the majority of I-E is matured and surface-expressed, which can be quantitatively assessed with radiolabeled 14-4-4S.

• Immunological Impact: By raising anti-idiotypic antibodies against 14-4-4S and using inhibition ELISA, researchers demonstrated that the 14-4-4S idiotype is a structurally shared idiotype, analogous to shared idiotypes seen in antibody responses to defined haptens and proteins. This was the first reported example of a shared monoclonal idiotype in the anti-MHC system.

• Experimental Controls and Purity: Purified antibody has high cytotoxic titers and shows a single band in immunoelectrophoresis, providing a robust reagent for immunological research.

These results establish 14-4-4S as a fundamental tool for dissecting MHC class II biology, idiotypic networks, and mechanisms of antigen presentation in murine models.

There is limited information in the search results regarding the specific dosing regimens of the clone 14-4-4S monoclonal antibody across different mouse models. However, the applications and usage of this antibody provide some insights into its general use.

The 14-4-4S monoclonal antibody is used to react with the mouse and rat major histocompatibility complex class II, specifically the I-E subregion-encoded glycoprotein, in mice strains of H-2^k, H-2^d, H-2^p, and H-2^r. It is typically used in flow cytometric analysis, immunoprecipitation, complement-dependent cytotoxicity, and immunohistochemical staining. For flow cytometric analysis, the recommended dose is less than or equal to 0.5 μg per test, and the antibody should be carefully titrated for optimal performance.

In terms of varying dosing regimens across different models, general practices in immunology can suggest that dosing may need to be adjusted based on the specific requirements of each model, such as the strain of mice, the nature of the experiment, and the specific application of the antibody. However, specific details on adjusting dosages for clone 14-4-4S across different mouse models are not provided in the search results.

For a more precise answer, consulting scientific literature specific to the use of clone 14-4-4S in different mouse models would be necessary.

General Considerations for Dosing Regimens

• Strain and Model Specificity: Different mouse strains may require different dosing due to variations in immune response and antibody clearance.
• Experimental Goals: The purpose of the experiment (e.g., depletion, blocking, or analysis) can influence the dosing strategy.
• Antibody Formulation: The form of the antibody, such as conjugation or fragment, can affect its pharmacokinetics and dosing requirements.

Due to the lack of specific information on clone 14-4-4S dosing regimens across different models, empirical titration and literature review are recommended for precise dosing strategies.