Anti-Mouse CD11c [Clone N418] — Purified in vivo PLATINUM™ Functional Grade

The clone N418 is widely used in in vivo mouse studies for targeting dendritic cells, which are crucial components of the immune system. This antibody specifically binds to CD11c, a marker predominantly expressed on dendritic cells in mice. Here are some key ways N418 is utilized:

1. Dendritic Cell Depletion: N418 is used to deplete dendritic cells in vivo, providing valuable insights into immune mechanisms and disease models by manipulating the role of these cells in immune responses.

2. Enhancing Antigen-Specific Responses: N418 has been reported to enhance antigen-specific responses when used to target dendritic cells in vivo. This is beneficial for studying immune responses and vaccine efficacy.

3. Immune System Studies: By targeting CD11c+ cells, researchers can investigate the function of dendritic cells in various immune contexts, such as inflammation, tolerance, and autoimmune diseases.

4. Applications: N418 is utilized in various applications, including ELISA, flow cytometry, and neutralization assays, which help in studying biological pathways affected by CD11c expression.

Overall, the N418 clone is a valuable tool for in vivo studies targeting dendritic cells in mice, offering insights into immune function and disease mechanisms.

The recommended storage temperature for sterile packaged clone N418 (anti-mouse CD11c monoclonal antibody) is -20°C to -70°C for long-term storage, and 2°C to 8°C for short-term storage (up to 1 month). Upon receipt, it should be immediately stored at one of these temperatures depending on your intended duration of use.

• For long-term stability (up to 12 months), store at -20°C to -70°C, using a manual defrost freezer and avoiding repeated freeze-thaw cycles.
• For short-term use (up to 1 month), storage at 2°C to 8°C is acceptable.
• Avoid storing in frost-free freezers and minimize freeze-thaw cycles, as these conditions may denature the antibody.

These guidelines ensure preservation of antibody activity and sterility for research applications. Always check the specific product datasheet with your supplier, as formulations and packaging may vary.

Based on the research literature, several antibodies and proteins are commonly used alongside the N418 anti-CD11c antibody in experimental studies.

Control and Comparison Antibodies

Hamster anti-TNP antibody serves as an important control in dendritic cell targeting experiments. Studies have shown that when hamster anti-TNP and goat anti-hamster IgG antibodies are injected separately, they do not result in an amplified immune response, unlike the specific interaction seen with N418. This demonstrates the specificity required for dendritic cell targeting.

J1.2 rat antibody has been used as a non-interacting control antibody in comparative studies with N418. Research has demonstrated that when mice are immunized with a single dose of N418 and the non-interacting J1.2 rat antibody, no rat-specific antibody response is detectable in mouse serum, further confirming the requirement for specific antigen-antibody interactions.

Target Antigens and Reporter Proteins

Polyclonal goat anti-hamster antibody is frequently employed as a model antigen in N418-based dendritic cell targeting studies. When this goat antibody is complexed with N418, it elicits robust goat antibody-specific serum IgG responses in mice, with detectable titers by day 5 and ranging from 1:1000 to 1:100,000 by day 7.

Goat anti-biotin antibody serves as another control antigen that demonstrates the specificity requirements of N418-mediated targeting. Research has shown that there is little to no interaction between goat anti-biotin antibody and N418, and the presence of N418 does not amplify the antibody response to this antigen - in fact, it may have a slightly suppressive effect in certain mouse strains.

Co-staining and Multi-parameter Analysis

CD3 antibodies are commonly used in conjunction with N418 for multi-parameter flow cytometry analysis. Studies routinely employ CD3 PE-conjugated antibodies alongside N418 to distinguish different cell populations, particularly when analyzing splenic cells and characterizing dendritic cell subsets.

CD45 and CD11b antibodies are frequently used in gating strategies with N418. Flow cytometry protocols often involve gating against CD45+ CD11b+ GR1- cells when using N418 to identify specific dendritic cell populations in tissues such as alveolar cells from bronchoalveolar lavage samples.

These combinations demonstrate that N418 is most effective when used with specific interacting antigens for targeting purposes, while non-specific or non-interacting antibodies serve as important controls to validate the specificity of dendritic cell-mediated immune responses.

Clone N418 is a monoclonal antibody widely used to detect murine CD11c, a key marker on dendritic cells, and has been cited extensively for its role in immunology, particularly flow cytometry and functional studies in mice. Key findings in scientific literature using clone N418 include:

• Identification and Isolation of Mouse Dendritic Cells: N418 is a standard tool for identifying CD11c+ cells (predominantly dendritic cells) in murine tissues, enabling immunophenotyping via flow cytometry and other platforms.

• Immunomodulation and Functional Enhancement: Clone N418 has been reported to enhance antigen-specific responses when used to target dendritic cells in vivo, suggesting a utility not just for identification but also for modulating immune activity in experimental settings.

• Assessment of Immune Suppression: In studies investigating the effects of stress on the immune system, clone N418 has been used to delineate dendritic cell responses and phenotypes, for example, showing how psychological stress affects dendritic cell populations and immune suppression.

• Routine Flow Cytometry Control: Detailed protocols using N418 as a positive marker for dendritic cells have been described, including gating strategies to exclude T and B cells and reliably assess MHC-II and CD11c expression in complex leukocyte mixtures.

• Breadth of Use and Standardization: The antibody is heavily cited across immunology research, making it a standard for murine dendritic cell work, especially in studies involving the analysis of immune subsets, cell activation states, and responses to vaccines, infections, or immune modulation.

Essential technical facts:

• Host: Armenian hamster monoclonal.
• Target: Mouse CD11c (Integrin alpha-X/Itgax).
• Applications: Primarily flow cytometry, also used in in vivo functional modulation.
• Enhances antigen-specific immune responses when targeting dendritic cells in vivo.

These findings have collectively established N418 as a gold standard reagent in mouse immunology for both basic and preclinical studies.