Anti-Mouse CD226 [Clone 10E5] – Purified in vivo PLATINUM™ Functional Grade

Clone 10E5 is a monoclonal antibody targeting mouse CD226 (DNAM-1) that is commonly used in vivo to functionally inhibit CD226 activity, particularly in studies of immune cell function and immune-mediated diseases in mouse models.

Key in vivo applications include:

• Blocking CD226-mediated T cell activity: 10E5 inhibits antigen-specific T cell proliferation and interferon-gamma (IFN-γ) production, making it useful for dissecting the role of CD226 in T cell responses and adaptive immunity.
• Studying autoimmunity: Administration of clone 10E5 in vivo can delay onset and reduce severity of Th1-mediated autoimmune diseases (e.g., models of multiple sclerosis, type 1 diabetes), highlighting its use in evaluating CD226’s role in autoimmune pathogenesis.
• Analyzing NK cell function: Since CD226 is also expressed on NK cells, 10E5 can be used to assess the contribution of CD226 to natural killer cell cytotoxicity and cytokine secretion.
• Examining broader immune responses: By inhibiting CD226, researchers explore its impact on intercellular adhesion, lymphocyte signaling, and the interplay between innate and adaptive immunity.

Summary table:

CD226 (DNAM-1) is broadly expressed on T cells (including subsets of thymocytes), NK cells, some B cells, monocytes, and platelets, making clone 10E5 a versatile tool for functional blockade in immune cell studies. In all cases, the antibody is frequently used at low doses and may be leveraged in both flow cytometric and functional in vivo assays.

The 10E5 antibody designation is used for several different monoclonal antibodies targeting distinct antigens, and the commonly co-used antibodies vary depending on the specific research context.

Platelet and Integrin Research

In studies involving the 10E5 antibody that targets platelet glycoproteins IIb/IIIa (αIIbβ3 integrin), several antibodies and proteins are frequently used alongside it. Anti-CD61 (β3 integrin) is commonly employed in integrin and platelet research. Natural ligands such as fibrinogen and von Willebrand factor (vWF) are also standard companions, as these proteins naturally interact with the integrin complex that 10E5 targets.

Additional commonly used antibodies in this context include activation-specific integrin antibodies like PAC-1, glycoprotein markers such as anti-CD42b, and appropriate isotype controls. Other monoclonal antibodies used in platelet research include 7E3 (anti-β3), AP3 (anti-β3), and AP2 (anti-αIIb/β3). The 10E5 antibody in this application has been shown to block fibrinogen binding to platelets and inhibit platelet aggregation induced by various agonists.

T Cell and Immunology Research

When 10E5 refers to the anti-CD226 (DNAM-1) antibody used in immunological studies, it is commonly employed in flow cytometric analysis of CD3/CD28-stimulated mouse splenocytes. In this context, the antibody has been reported to inhibit antigen-specific T cell proliferation and IFN-gamma production. CD226/DNAM-1 is broadly expressed on T cells, NK cells, platelets, monocytes, and a subset of B cells.

p53 Research

The 10E5 clone also exists as an antibody targeting p53 acetylated at lysine 120. In this application, it is used alongside antibodies such as anti-Myc, anti-His-tag, and general anti-p53 antibodies like DO-1 or FL393 for immunoprecipitation and Western blotting studies.

Clone 10E5 is a designation used for multiple monoclonal antibodies in scientific research, most notably as:

• An anti-human integrin αIIb (GPIIb) antibody that blocks fibrinogen binding and platelet aggregation
• An anti-mouse CD226 (DNAM-1) antibody that regulates T cell and NK cell function
• An anti-human CD9 and anti-human p53 acetyl-K120 antibody in specific contexts

Key findings from scientific literature citing clone 10E5, grouped by target/context:

1. Anti-human Integrin αIIb (GPIIb) Clone 10E5

• Binds to the cap subdomain of GPIIb: Crystallographic and immunochemical studies pinpoint that 10E5 targets the "cap" of the αIIb β-propeller domain, directly interacting with repeats 1–3.
• Blocks ligand binding: 10E5 inhibits fibrinogen and von Willebrand factor binding to αIIbβ3, effectively preventing platelet aggregation in normal platelets.
• Useful for epitope mapping and structure-function studies: Extensively used to characterize conformational changes and binding sites on GPIIb/IIIa, as its binding is altered by N-terminal substitutions in the β-propeller domain.
• Does not bind Glanzmann thrombasthenia platelets: 10E5 fails to react with platelets lacking functional αIIbβ3, making it a diagnostic tool for this disorder.

2. Anti-mouse CD226 (DNAM-1) Clone 10E5

• Widely used in flow cytometry and in vivo inhibition studies: Frequently cited for detection and functional blockade of CD226 on T cells, NK cells, monocytes, and platelets.
• Modulates T cell and NK cell responses:
  • Inhibits antigen-specific T cell proliferation and IFN-γ production in vivo.
  • Used in studies demonstrating the role of CD226 in lymphocyte adhesion, activation, cytotoxic function, and cytokine secretion.
• Functional-grade antibody: Reported as a reliable functional blocking and depletion reagent in murine models.

3. Other Contexts for "10E5" (Less Common)

• Anti-CD9 (Human B cells): 10E5 was used to label CD9+ germinal center B cells, identifying plasma cell precursors in human tonsil tissue.
• Anti-p53 (acetyl K120) in human studies: Cited for detecting acetylated p53 at lysine 120 in various human cell analyses.

Summary Table: Primary Targets and Key Findings for Clone 10E5

Conclusion:
Scientific literature referencing clone 10E5 highlights its use as a highly specific and functionally potent monoclonal antibody in diverse research fields—most notably for blocking platelet integrin function and for immunophenotyping or functional blocking of CD226 in murine immunology. Its specificity and blocking capabilities make it a preferred tool in mechanistic studies of platelet aggregation and immune cell regulation.

Dosing regimens for clone 10E5 (anti-mouse CD226) vary significantly depending on the specific experimental goals and mouse models being used, but there is no universally fixed regimen established in the literature. The optimal dose and frequency must be carefully titrated for each specific application.

Factors Influencing Dosing Variation

The variation in dosing regimens across different mouse models is influenced by several key factors. Mouse strain differences play a critical role, as different strains may require adjusted tamoxifen doses and injection routes for specific applications. Additionally, the route of elimination, metabolism, and half-life of antibodies can differ substantially and may even vary between different genetic backgrounds, which accounts for varying dosing requirements.

General Considerations for Optimization

When establishing a dosing regimen for clone 10E5 in a particular mouse model, researchers should consider that careful titration is essential for optimal performance in the specific assay of interest. This optimization process typically involves testing different doses and schedules to achieve the desired level of target engagement and functional outcome.

Context from Related Antibodies

While specific dosing information for clone 10E5 is limited, examining other immune checkpoint and cell marker antibodies provides useful context. For instance, checkpoint blockade antibodies like anti-PD-1 are commonly used at 200-500 μg per mouse with dosing every 3-4 days, while cell-depleting antibodies such as anti-CD4 (GK1.5) are typically administered at 200-250 μg per mouse 2-3 times per week. These examples illustrate that dosing regimens are highly dependent on the antibody's mechanism of action and the desired biological effect.

The lack of standardized dosing protocols for clone 10E5 underscores the importance of pilot studies to determine optimal parameters for each experimental system.