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

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

Product No.: C2482

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Clone
10E5
Target
CD226
Formats AvailableView All
Product Type
Hybridoma Monoclonal Antibody
Alternate Names
DNAM-1, PTA1 (platelet and T cell activation antigen 1), TLISA1, LFA-1 associated Molecule PTA-1
Isotype
Rat IgG2b κ
Applications
B
,
FA
,
FC
,
in vivo

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Dilution Buffer
Immunogen
Murine Th1 cell clone AE7 and Th1 cell lines derived from 5B6 and DO11.10 TCR transgenic mice
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<0.5 EU/mg as determined by the LAL method
Purity
≥98% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
State of Matter
Liquid
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM<sup>TM</sup> antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Regulatory Status
Research Use Only
Country of Origin
USA
Shipping
2 – 8° C Wet Ice
Additional Applications Reported In Literature ?
B,
FA,
FC,
in vivo
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
10E5 activity is directed against murine CD226 (DNAM-1; PTA-1).
Background
Murine (m) CD226 (DNAM-1; PTA1) is an adhesion molecule involved in intercellular adhesion, lymphocyte signaling, cytotoxicity, and lymphokine secretion mediated by cytotoxic T- lymphocyte and natural killer (NK) cells (Uniprot Q8K4F0)1. mCD226 is also essential for NK- mediated killing of immature dendritic cells 2 and acts as an important costimulatory molecule in Th1 cell activation and effector function3.

CD226 binds to PVR (CD155) via its N-terminal Ig-like domain2 and with cell surface receptor NECTIN2 (CD112)1. Based on its similarity to human CD226 (Uniprot Q15762), mCD226 likely competes with PVRIG for NECTIN2 binding.

mCD226 is a marker for mature T cells2 and is specifically expressed on the surface of differentiated Th1 cells but not Th2 or Th0 cells3. mCD226 expression is enhanced by Th1 differentiation but down-regulated upon Th2 polarization.

10E5 was generated by immunizing Lewis and Lou/M rats with established murine Th1 cell clone AE7 and Th1 cell lines derived from 5B6 and DO11.10 TCR transgenic mice3. The resulting panel was screened on polarized Th1 and Th2 T cell clones and cell lines to find an antibody that selectively recognizes only Th1 clones. 10E5 specifically recognizes CD226 on the surface of Th1 cells. Additionally, 10E5 inhibits Ag-specific T cell expansion, reducing the expansion of PLP 139-151-specific Th1 cells producing IFN-γ. In vivo treatment with anti- CD226 clone 10E5 inhibits T cell activation and delays the onset and reduces the severity of Th1-mediated autoimmune disease.

Antigen Distribution
mCD226 is expressed mainly on T cells, including on the surface of differentiated Th1 cells, almost all naïve CD8+ T cells, activated CD4+ cells, about 40% of unactivated CD4+ T cells as well as on subsets of primary CD11b+ macrophages, CD11c+ dendritic cells and NK1.1 and DX5 NK cells.
Ligand/Receptor
CD112 and CD155
NCBI Gene Bank ID
UniProt.org
Research Area
Immunology
.
Adhesion Molecules
.
CD Molecules

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

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:

Application ContextFunctional Outcome of 10E5 In Vivo Use
T cell inhibitionBlocks antigen-specific proliferation and IFN-γ production
Autoimmunity modelsDelays onset, reduces severity of Th1-driven disease
NK cell studiesUsed to examine role of CD226 in cytotoxicity/cytokine production
Immune signalingDissects CD226’s involvement in lymphocyte interactions

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

TargetKey ApplicationPrincipal Findings & Uses
Integrin αIIbGPIIb/IIIa Platelet StudiesBlocks fibrinogen binding, epitope mapping, diagnostic for Glanzmann thrombasthenia
CD226 (DNAM-1)Murine ImmunologyInhibits T cell/NK function, flow cytometry, functional blockade in vivo
CD9Human ImmunologyMarker for plasma cell precursors in B cell differentiation studies
p53 (acetyl-K120)Human Cancer/Cell BiologyDetects acetylated p53, used in cell cycle and apoptosis research

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.

References & Citations

1. https://www.uniprot.org/uniprotkb/Q8K4F0/entry
2. Seth S, Georgoudaki AM, Chambers BJ, et al. J Leukoc Biol. 86(1):91-101. 2009.
3. Dardalhon V, Schubart AS, et al. J Immunol. 175(3):1558-1565. 2005.
B
FA
Flow Cytometry
in vivo Protocol

Certificate of Analysis

Formats Available

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.