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

Clone 10E5 is primarily used in mice for in vivo functional inhibition of CD226 (DNAM-1), a receptor expressed on immune cells such as T cells and NK cells. The most common applications are:

• Inhibition of antigen-specific T cell proliferation: Blocking CD226 with clone 10E5 prevents the expansion of T cells in response to specific antigens.
• Suppression of interferon-gamma (IFN-γ) production: CD226 blockade leads to reduced IFN-γ secretion, affecting Th1 immune responses.
• Delayed onset and reduced severity of Th1-mediated autoimmune disease models: In vivo administration of clone 10E5 inhibits T cell activation and is reported to delay and reduce the severity in Th1-dominated autoimmune disease models, such as experimental autoimmune encephalomyelitis (EAE).
• Functional studies of DNAM-1 signaling: Used to study the role of DNAM-1 in cytotoxicity, lymphocyte signaling, and lymphokine secretion.

Additional details:

• Selectivity: Clone 10E5 binds specifically to differentiated Th1 cells, not Th0 or Th2 cells.
• Assay compatibility: While the antibody is also used for flow cytometry and cell staining, its major in vivo role is functional blockade.
• Mechanistic research: Used for dissecting pathways involving intercellular adhesion and immune regulation via CD226.

In summary, clone 10E5 is a tool for investigating and manipulating Th1-driven immune responses, autoimmune disease progression, and the functional biology of CD226 in live mouse models.

Several different monoclonal antibodies and proteins are commonly used in conjunction with 10E5, depending on the experimental model and research focus. The most frequent combinations in the literature are seen in platelet/integrin biology and immune cell signaling research.

For platelet and integrin αIIbβ3 (GPIIb/IIIa) studies (where 10E5 is an anti-αIIb monoclonal antibody):

• Anti-CD61 (β3 integrin): Used alongside 10E5 to characterize the αIIbβ3 complex, since CD61 identifies the β3 subunit.
• 7E3 (anti-β3 integrin): Another monoclonal that specifically binds β3, often used in comparison or co-staining with 10E5.
• AP3 and AP2 (anti-β3, anti-αIIb/β3): These recognize either the β3 integrin or the complete complex, providing additional epitope coverage in studies of platelet aggregation and fibrinogen receptor biology.
• Fibrinogen (as a protein ligand): Fibrinogen binding assays using 10E5 are standard, since 10E5 blocks fibrinogen interaction with platelets, allowing measurement of receptor-ligand interactions.

For CD226 (DNAM-1) studies (where 10E5 is an anti-CD226 antibody):

• CD3 and CD28 antibodies: Frequently used to stimulate T cell activation in studies evaluating the effect of CD226 blockade by 10E5 on T cell proliferation and cytokine secretion.
• Markers for T cell, NK cell, monocyte, and B cell subsets: These markers are often combined with 10E5 for flow cytometric phenotyping, including antibodies against CD3, CD4, CD8, CD56, and CD19.

Other notable usages:

• Anti-p53 antibodies: In research focusing on p53 acetylation, 10E5 has been used to detect p53 acetylated at Lys120, commonly alongside total p53 antibodies (e.g., DO1, FL393) and markers such as Myc or His-tag for protein overexpression validation.

Summary Table: Common Antibody/Protein Combinations with 10E5

These combinations are methodological standards for dissecting molecular interactions, cell surface phenotype, and downstream functional consequences in flow cytometry, immunoprecipitation, and Western blotting.

Clone 10E5 has been cited in scientific literature in two major contexts: as an antibody specific for integrin αIIb (CD41) on platelets and as an antibody targeting CD226 (DNAM-1) on lymphocytes. The key findings are:

• Integrin αIIb Antibody (10E5):

  • 10E5 binds specifically to the cap subdomain of the β propeller domain of integrin αIIb (GPIIb).
  • The antibody inhibits ligand binding (including fibrinogen and von Willebrand factor) to the integrin αIIbβ3 complex, thereby blocking platelet aggregation.
  • Crystallographic studies have mapped its epitope to contacts involving the first three repeats of the cap subdomain.
  • 10E5 does not react with platelets from patients with Glanzmann thrombasthenia, confirming its specificity for functional αIIb.
  • It does not work for Western blotting applications but is extensively used for flow cytometry and immunoprecipitation.

• Anti-CD226 (DNAM-1) Antibody (10E5):

  • Clone 10E5 recognizes mouse CD226 (DNAM-1), a receptor involved in lymphocyte signaling, cytotoxicity, and intercellular adhesion.
  • It is routinely used for flow cytometric analysis and has been reported to effectively inhibit antigen-specific T cell proliferation and IFN-γ production in vivo.
  • DNAM-1 is broadly expressed on T cells, NK cells, platelets, monocytes, and some B cells, underscoring the antibody's utility in immunological studies.
  • The clone is well-documented for use in immune suppression experiments and functional characterization of cytotoxic lymphocytes.

• Other applications and citations:

  • Clone 10E5 has also been used for identifying CD9 expression on plasma cell precursors as well as for staining in flow cytometry in multiple hematological and immunological studies, sometimes with anti-CD9 and anti-CD226 together.
  • There is a similarly named antibody (10E5) directed against p53 acetyl K120, but this is likely a different clone, based on the distinct epitope and application (detecting posttranslational modification of p53).

Summary Table:

These citations establish 10E5 as a well-characterized monoclonal antibody with broad utility in both platelet biology (integrin αIIb studies) and immunology (CD226 functional profiling). The evidence comes from lineage-tracing, crystallographic mapping, ligand inhibition studies, and standardized flow cytometry protocols. Discrepancies or alternate uses likely reflect distinct clones sharing the 10E5 name, so cross-referencing the target antigen is essential for interpretation.

Dosing regimens of clone 10E5 (anti-mouse CD226/DNAM-1) vary considerably based on the mouse model and the specific experimental objectives, and there is no universally established protocol. This variability is noted by reagent providers, which emphasize that dosing must be tailored to the study design, the mouse strain, and endpoints such as T cell activation inhibition, tumor growth, or other immunological parameters.

Key considerations for dosing clone 10E5 include:

• Experimental goals: Studies may focus on different outcomes (e.g., T cell activation, tumor rejection), influencing both dose and schedule.
• Mouse strain/model: Immunological status (e.g., wild-type, knockout, humanized) can affect the optimal dose and frequency.
• Dosing guidance: Product technical sheets recommend titrating clone 10E5 for optimal performance in each assay or model, rather than adhering strictly to a published standard.
• General antibody dosing in mice: For similar functional-grade antibodies, common regimens often range from 100–250 μg per mouse via intraperitoneal injection, 2–3 times per week, although specific studies with 10E5 may adjust this further depending on tolerability and effect.

Example guidance and recommendations:

• Leinco Technologies notes, “Dosing regimens for clone 10E5 … vary depending on experimental goals and mouse models, but there is no universally fixed regimen; dose and frequency should be empirically optimized”.
• Thermo Fisher states, “It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest”.

In summary:
There is no single standard dosing regimen for clone 10E5 across all mouse models. Instead, researchers are advised to determine the dose empirically, typically beginning with protocols used for similar T cell or immune checkpoint antibodies, and then refining based on pilot results and study-specific needs. Published studies using this clone may report further model-specific regimens, but such protocols remain variable and context-dependent.