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

Clone 10E5 is a rat anti-mouse monoclonal antibody that targets CD226 (DNAM-1) and is commonly used in in vivo mouse studies to functionally block or inhibit CD226 receptor activity on immune cells.

The principal uses and mechanisms in in vivo mouse studies include:

• Functional Inhibition of CD226: 10E5 is primarily used to block the functional activity of CD226 on mouse immune cells. In vivo, this blockade has been reported to inhibit antigen-specific T cell proliferation and interferon-gamma (IFN-?) production. This suggests its utility in studies dissecting the role of CD226 in T cell activation, immune regulation, and autoimmune or tumor models.

• Experimental Design: In in vivo experiments, mice are typically administered the 10E5 antibody systemically (e.g., via intraperitoneal or intravenous injection) to achieve blockade of CD226 function on relevant immune cell populations, such as T cells and NK cells. The exact dosing and schedule depend on the experimental protocol and the biological question. The antibody can be combined with other immune-modulating agents to study synergistic or antagonistic effects.

• Readouts and Applications: Inhibition mediated by clone 10E5 in vivo is monitored by:

  • Assessing suppression of T cell proliferation,
  • Decreased production of IFN-? (a key cytokine in immune responses),
  • Studying immune regulation in the context of tumor immunology, infection, or autoimmunity.

• Validation and Controls: Clone 10E5 is also used in flow cytometry to detect CD226 expression on mouse immune cells, and its blocking effect can be confirmed by loss or reduction of CD226-mediated responses.

• Supporting Literature: Published studies have described use of 10E5 to specifically inhibit T cell expansion and to examine antigen-presenting cell (APC) function in vivo, indicating its robust utility across immunological assays.

In summary, clone 10E5 is a key tool antibody for in vivo mouse studies aimed at dissecting the functional role of CD226 in immune responses by specifically blocking its activity, thereby enabling detailed mechanistic studies of T cell and NK cell biology in health and disease.

The correct storage temperature for sterile packaged monoclonal antibody clones, such as clone 10E5, depends on the intended duration and conditions of storage:

• For short-term storage (up to one month), store at 2–8°C in a refrigerator.
• For long-term storage (months to years), aliquot and store at ? -20°C, and ideally at ? -70°C or -80°C for maximum stability.

Additional Details:

• Avoid repeated freeze-thaw cycles: These can denature the antibody and reduce activity. Aliquot into working volumes to minimize this risk.
• Do not store at room temperature for extended periods; monoclonal antibodies may experience activity loss if not refrigerated or frozen.
• Store in the back of the fridge or freezer to avoid temperature fluctuations, which are more common near doors.

If the antibody is for therapeutic use or under specific manufacturer instructions, check the product datasheet for any unique requirements. The above guidelines cover typical research and functional grade monoclonal antibodies like clone 10E5.

The monoclonal antibody 10E5 is most commonly used to detect the native conformation of integrin alphaIIb (CD41) in complex with beta3 (CD61), making it highly relevant in studies focused on the platelet GPIIb/IIIa integrin complex. Literature featuring 10E5 often pairs it with the following antibodies or proteins, depending on the research context:

• Anti-beta3 (CD61) antibodies: Since 10E5 binds the alphaIIb subunit of integrin ?IIb?3, researchers frequently use antibodies against the beta3 subunit to study the complete receptor complex or to confirm co-localization and complex formation.
• Fibrinogen and von Willebrand factor (vWF): These are the physiological ligands for integrin ?IIb?3, and are often used in functional assays together with 10E5 to assess ligand binding, platelet aggregation, or to quantify antibody inhibition of these interactions.
• Activation-specific integrin antibodies (e.g., PAC-1): PAC-1 recognizes the activated form of integrin ?IIb?3, so it is often used alongside 10E5 to distinguish between integrin activation states in flow cytometry and functional studies.
• Other platelet glycoprotein antibodies (e.g., CD42b for GPIb, anti-P-selectin/CD62P): To profile platelets or establish lineage specificity, studies may include additional markers for multi-parameter flow cytometry or immunoprecipitation.
• Secondary anti-mouse IgG antibodies: Because 10E5 is a mouse IgG2a monoclonal antibody, species-specific secondary antibodies are used for detection in IHC, IF, FACS, or ELISA experiments.

When 10E5 is used for functional blocking, it is frequently assessed in parallel with these controls:

• Isotype-matched control antibodies: To account for non-specific binding or effects in blocking or flow assays.
• Alternative integrin-blocking antibodies (e.g., 7E3, LM609, or ReoPro/abciximab): Different anti-?IIb?3 antibodies may be used to compare blocking efficacies and to map epitope specificities.

Note: There are additional unrelated "10E5" clones targeting molecules such as p53 (Acetyl Lys120), IL-6, or CD226/DNAM-1 in other contexts. Cross-check the 10E5 clone application to ensure it refers to the integrin ?IIb antibody.

In summary, the antibodies and proteins most commonly used with 10E5 in integrin and platelet research are anti-CD61 (?3 integrin), natural ligands (fibrinogen, vWF), activation-specific integrin antibodies (like PAC-1), glycoprotein markers (such as anti-CD42b), and isotype controls.

Key Findings from Clone 10E5 Citations in Scientific Literature

Clone 10E5 is referenced in two distinct scientific contexts: as an anti-mouse CD226 antibody and as an anti-human integrin ?IIb antibody. The key findings for each are summarized separately below.

Anti-Mouse CD226 (Clone 10E5)

• Generation and Origin: Clone 10E5 is a monoclonal antibody generated by immunizing Lewis and Lou/M rats with established murine Th1 cell clones (AE7) and cell lines (5B6 and DO11.10 TCR cells), specifically targeting CD226.
• Primary Use: While detailed findings on the functional consequences of using this clone (e.g., in biological assays or animal studies) are not provided in the available literature, clone 10E5 has been used as a reagent for flow cytometry, as indicated by its inclusion in panels for murine immune cell characterization.
• Available Information: The available citation highlights the clone’s production method but does not elaborate on mechanistic or functional studies in scientific reports. Further details about its in vivo or in vitro effects remain to be explored in more detailed publications.

Anti-Human Integrin ?IIb (Clone 10E5)

• Clone Characteristics: 10E5 is a murine monoclonal antibody (isotype MIgG2a) that recognizes the human integrin ?IIb subunit, specifically in its native conformation when associated with the ?3 subunit.
• Functional Properties: The antibody is a potent inhibitor of ligand binding (fibrinogen and von Willebrand factor) to the integrin ?IIb?3 complex and blocks platelet aggregation. Critically, it does not react with platelets from patients with Glanzmann thrombasthenia—a disease characterized by defective ?IIb?3 expression—confirming its specificity for the intact receptor.
• Epitope Mapping: The epitope is located in the ?IIb cap subdomain, further defining the molecular interaction site.
• Applications: Recommended for ELISA, FACS, function blocking, and immunoprecipitation, but not for Western blotting, suggesting it is conformation-dependent and may not detect denatured protein.
• Scientific Impact: This clone has been widely used in hematology research to dissect the role of ?IIb?3 in platelet function, thrombosis, and bleeding disorders, influencing both basic science and clinical diagnostics.

Summary Table

In summary, clone 10E5 is most prominently recognized for its role as a function-blocking antibody against human integrin ?IIb, with clear findings on its specificity, functional inhibition, and clinical relevance in platelet biology. Its use as an anti-mouse CD226 clone is documented, but the available literature provides only limited details on its effects or scientific findings. For comprehensive mechanistic insights, especially for anti-CD226 10E5, further primary research articles would be required.