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

Clone 10E5 is a rat anti-mouse monoclonal antibody targeting CD226 (DNAM-1), and it is used in mouse in vivo studies primarily to inhibit T cell activation and modulate immune responses, especially those involving Th1-mediated autoimmune processes. In experimental models, 10E5 is administered to mice to:

• Inhibit antigen-specific T cell expansion and IFN-? production: Treatment with 10E5 dampens the proliferation of CD4+ Th1 cells reactive to specific antigens and reduces their cytokine secretion (e.g., IFN-?), important in autoimmune disease models.
• Delay onset and reduce severity of Th1-mediated autoimmune disease: By blocking CD226 function in vivo, 10E5 can attenuate T cell activation, thereby delaying and decreasing the severity of autoimmune conditions that depend on Th1 cell responses.
• Functional immunological assays and flow cytometry: 10E5 is also employed to measure CD226 expression on various immune subsets and to functionally block CD226 in vitro and in vivo experiments, including analyses of T cell, NK cell, and other leukocyte populations.

Mechanism and Cellular Targets:

• CD226 (DNAM-1) is expressed predominantly on T cells (especially naïve CD8+ and activated CD4+), Th1 cells, NK cells, and some myeloid and B cell subsets.
• Interfering with CD226 impacts lymphocyte adhesion, signaling, cytotoxicity, and differentiation, all of which are critical for adaptive and innate immunity.

Typical in vivo applications:

• Administration of purified functional-grade clone 10E5 antibody to live mice to block CD226, often as part of studies investigating immune cell activation, autoimmune disease pathogenesis, or immune modulation by targeting Th1 cell responses.
• Used in flow cytometry and functional blockade experiments to study immunological mechanisms and cell population dynamics.

Key references from the literature confirm that anti-CD226 (clone 10E5) is a well-established experimental tool for functional blockade in mouse models, particularly for studying Th1-mediated autoimmunity and T cell biology.

The 10E5 antibody is most commonly used as an inhibitor or blocking antibody targeting different proteins depending on context. However, the classic and most frequently referenced usage in the literature is anti-integrin ?IIb (CD41) on platelets for the study of the glycoprotein IIb/IIIa complex (also known as integrin ?IIb?3). When used in such experiments, 10E5 is often combined with several other antibodies and proteins to dissect platelet function and integrin biology.

Commonly used antibodies or proteins with 10E5 in the literature:

• Anti-GPIIb (integrin ?IIb, CD41) and anti-GPIIIa (integrin ?3, CD61) antibodies: These are directly associated with the IIb/IIIa complex and often co-used to confirm specificity or for dual/inhibition studies.
• Fibrinogen: Since 10E5 blocks fibrinogen binding to IIb/IIIa, exogenous fibrinogen is frequently added as a ligand to induce platelet aggregation, and the effects of 10E5 are compared to other inhibitors or proteins affecting this interaction.
• Agonists such as ADP, epinephrine, or thrombin: These are commonly used to stimulate platelets, with and without 10E5, to study activation and aggregation pathways.
• Chymotrypsin-treated and aspirin-treated platelets: As experimental controls or to modify the state of the GPIIb/IIIa complex and test the function/blocking by 10E5.
• Other isotype control antibodies and function-blocking antibodies: To ensure specificity and for comparative inhibition studies (exact clones vary depending on the experiment).

Alternate Uses of "10E5" in Literature:

• Anti-mouse CD226 (DNAM-1) antibody: In some immunology contexts, 10E5 refers to a clone used against CD226—often co-used with other T-cell and NK-cell markers (e.g., anti-CD4, anti-Th1/Th2 differentiation markers).
• Anti-p53 (Acetyl Lys120) antibody: In molecular biology, a 10E5 clone targets acetylated p53 and is used alongside anti-Myc, anti-His, and total p53 antibodies in immunoprecipitation and western blotting settings.

Summary Table: 10E5 Contexts and Commonly Co-used Reagents

Conclusion:
The most common companions for 10E5 in its canonical platelet biology context are anti-GPIIb/IIIa antibodies and functional ligands like fibrinogen; in p53 and immunology contexts, it is paired with antibodies relevant to the respective pathways or cell subtypes. Always confirm which 10E5 clone and target are in use, as the clone designation overlaps across several different applications in the literature.

Clone 10E5 is primarily cited for its role as a monoclonal antibody used in immunological research to recognize specific cell surface proteins, with two distinct usages highlighted in the literature: anti-mouse CD226 and anti-human integrin ?IIb.

Key findings from scientific literature citing clone 10E5 include:

• Anti-mouse CD226 (DNAM-1):

  • Specificity: 10E5 targets murine CD226, a critical adhesion molecule involved in intercellular signaling, lymphocyte activation, cytotoxicity, and lymphokine secretion, especially among cytotoxic T lymphocytes and natural killer (NK) cells.
  • Selective Th1 Recognition: 10E5 was developed to recognize CD226 on Th1 cells, but not Th2 or Th0 cells. This makes it useful in dissecting Th1-mediated immune responses.
  • Functional Blocking: In vivo, anti-CD226 clone 10E5 inhibits antigen-specific T cell expansion (notably PLP 139-151-specific Th1 cells producing IFN-?), delays onset, and reduces severity of Th1-mediated autoimmune disease. This highlights its application in autoimmune and immunopathology models.
  • Expression Profile: CD226 identified by 10E5 is mainly expressed on differentiated Th1 cells, naïve CD8+ T cells, activated CD4+ T cells (about 40% of unactivated CD4+ T cells), as well as primary macrophage, dendritic, and NK subsets.
  • CD226 Knockout Effects: Publications using 10E5 as a research tool indicate that CD226 knockout results in protection against high-fat diet-induced obesity and systemic inflammation through impaired M1 macrophage polarization.
  • Citation: Dardalhon V, Schubart AS, et al. J Immunol. 175(3):1558-1565, 2005 is a pivotal paper using 10E5 for Th1 cell characterization.

• Anti-human integrin ?IIb (CD41):

  • Epitope & Function: 10E5 recognizes the ?IIb (integrin ?IIb) cap subdomain in its native conformation associated with ?3 integrin, inhibiting binding of fibrinogen and von Willebrand factor and thus blocking platelet aggregation.
  • Clinical Subsets: The antibody does not react with platelets from patients with Glanzmann thrombasthenia, confirming both functional specificity and diagnostic value.
  • Applications: Used for ELISA, flow cytometry, functional blocking, and immunoprecipitation. It is not suitable for Western blotting.

Summary Table — Clone 10E5 Antigen Targets and Major Findings

Additional Notes:

• 10E5 is widely cited for its functional blocking capacity and specificity in cell biology and immunology, enabling mechanistic studies in T cell and platelet biology.
• For all research using 10E5, material contributors and repositories like the Developmental Studies Hybridoma Bank (DSHB) should be cited for reagent use.

If your interest is in a specific clone 10E5 application or citation context (e.g., CD226 vs integrin ?IIb studies), please clarify for a more targeted synthesis.

Dosing regimens for clone 10E5 (anti-CD226, DNAM-1) in mouse models are not standardized in the literature, and the manufacturer (Thermo Fisher) notes that the antibody should be "carefully titrated for optimal performance in the assay of interest" due to potential variability based on application and experimental system. This means dosing can vary significantly depending on experimental purpose (e.g., flow cytometry, in vivo blocking, or functional inhibition), mouse strain, disease model, and target cell frequency.

Key details:

• Clone 10E5 targets CD226 (DNAM-1), a molecule on T cells, NK cells, and other immune cells, with expression levels and roles varying by cell subset and activation state.
• Intended use: 10E5 has been reported for applications like flow cytometry and has also been used in vivo to inhibit T cell proliferation and IFN-? production, implying possible use as a blocking or depleting antibody.
• Dosing instructions: No universal dosing regimen is provided; users are advised to titrate for the specific assay and mouse model. This is in contrast to several well-characterized clones such as RMP1-14 (anti-PD-1) or 2.43 (anti-CD8), for which standard in vivo doses (e.g., 200–250 ?g/mouse, intraperitoneally, 2–3 times/week) are common.

In summary:

• Dosing for 10E5 is highly context-dependent. Published literature or antibody datasheets do not specify standard in vivo dosing regimens as with common checkpoint or depletion antibodies.
• Optimization for each setting and mouse model is required, particularly for in vivo applications such as inhibition of CD226 function or immune modulation.
• For comparison, checkpoint blockade or immune cell–depleting antibodies in mouse studies are typically administered at 100–250 ?g/mouse intraperitoneally, 2–3 times per week. This range may serve as a starting point for pilot titrations with uncharacterized antibodies like 10E5, but requires empirical validation.

If you provide the exact application (e.g., blocking in tumor models, flow cytometry, depletion), a more precise estimate or precedent from the literature might be identified. Currently, no standard dose for 10E5 across models is published, and the dose must be empirically determined for each experimental use.