Anti-Mouse CD3ε [Clone 145-2C11] — Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse CD3ε [Clone 145-2C11] — Purified in vivo PLATINUM™ Functional Grade

Product No.: C2280

[product_table name="All Top" skus="C2280"]

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Clone
145-2C11
Target
CD3ε
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
CD3, T3
Isotype
Armenian Hamster IgG
Applications
Act
,
B
,
CyTOF®
,
Depletion
,
FA
,
FC
,
ICC
,
IF
,
IHC FF
,
in vivo
,
PhenoCycler®
,
WB

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Select Product Size
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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Armenian Hamster
Recommended Dilution Buffer
Immunogen
H-2Kb-specific mouse cytotoxic T lymphocyte clone BM10-37
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.
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™ 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.
Country of Origin
USA
Shipping
Next Day 2-8°C
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for this 145-2C11 antibody for staining cells in flow cytometry is ≤ 1.0 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application.
Additional Applications Reported In Literature ?
IHC (Frozen) The suggested concentration for this 145-2C11 antibody in IHC staining on frozen tissue is 5.0 - 10 μg per ml. Titration of the reagent is recommended for optimal performance for each application.
IP
Act
B
Depletion
ICC

CyTOF®
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
Clone 145-2C11 recognizes an epitope on mouse CD3ε.
Background
CD3ε is a 20kDa subunit of the TCR complex and is a transmembrane T-cell surface glycoprotein that belongs to the Ig superfamily. It is one of five polypeptide chains that form the TCR complex by associating with the CD3δ, γ and ζ chains, in addition to the TCR α/β or γ/δ chains. CD3 is involved in TCR signaling, enumeration of immunocompetent T-lymphocytes in peripheral blood, and signal transduction during antigen recognition.
Antigen Distribution
CD3ε is primarily expressed on mature T cells and NK-T cells and at different levels on differentiation-dependent thymocytes.
Ligand/Receptor
Peptide antigen/MHC-complex
PubMed
NCBI Gene Bank ID
Research Area
Immunology

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 145-2C11 is a monoclonal antibody targeting the mouse CD3? chain and is widely used in in vivo mouse studies primarily for T cell activation, depletion, and immunomodulation.

Key in vivo uses of 145-2C11 in mice include:

  • T cell Activation and Cytokine Release: Upon in vivo administration, 145-2C11 crosslinks CD3? on the surface of T cells, causing robust T cell activation, release of cytokines like TNF, IFN?, and IL-6, and often transient T cell receptor (TCR) downregulation. This property is exploited to model T cell activation or immune storm conditions in experimental animals.
  • In vivo T Cell Depletion: 145-2C11 can induce rapid depletion of peripheral T cells; this is widely used to study immune responses or generate T cell-deficient models. The antibody can induce apoptosis in immature thymocytes and deplete mature T cells in peripheral organs.
  • Immunosuppression: By triggering profound T cell modulation and depletion, 145-2C11 immunosuppresses mice, making it a surrogate for testing anti-CD3 therapies and mechanisms of tolerance induction.

Additional details and considerations:

  • Dosing and Monitoring: Studies have reported single or repeated intraperitoneal injections (e.g., 20?µg per mouse) causing acute T cell activation, rapid surface TCR downregulation, profound cytokine release, and temporary body weight loss (attributable to cytokine storm).
  • In vivo vs. in vitro utility: While 145-2C11 is also used in vitro for T cell activation and flow cytometry, in vivo its immunomodulatory and depleting effects predominate.
  • Use in Pathogenesis and Therapy Models: The antibody is used to mimic clinical anti-CD3 therapy, assess immune reconstitution, or study T cell-mediated pathologies.

Special preparation is required for in vivo applications (e.g., low endotoxin formulations such as Ultra-LEAF™) to prevent experimental confounding by contaminants.

In summary, 145-2C11 is most commonly used in vivo for T cell depletion, activation, and functional modulation in a range of immunological and disease models in mice.

The correct storage temperature for sterile packaged clone 145-2C11 (anti-mouse CD3? monoclonal antibody) is between 2°C and 8°C (refrigerator temperature), undiluted, and protected from prolonged exposure to light. Do not freeze the antibody.

This guideline applies to both conjugated and unconjugated forms of 145-2C11. Storing outside this range or freezing can damage the antibody, affecting its specificity and activity. Most suppliers recommend storage in the dark as some conjugates (like PE or APC) are light sensitive. If the antibody is supplied in solution, keep it undiluted unless the supplier’s protocol instructs otherwise.

Key points:

  • Store between 2°C and 8°C
  • Keep undiluted
  • Protect from light (if conjugated)
  • Do not freeze

In the literature, 145-2C11 (an anti-mouse CD3 antibody) is frequently used alongside several other antibodies and proteins to study T cell biology and immunomodulation in mice. Commonly co-used molecules include:

  • Anti-CD4 and Anti-CD8 Antibodies: These are routinely combined with 145-2C11 for immunophenotypic analyses of distinct T cell subsets by flow cytometry.
  • Concanavalin A (Con A): This lectin is utilized as an alternate T cell stimulator to assess T cell proliferative capacity in comparison or combination with anti-CD3 stimulation.
  • Methylprednisolone (Glucocorticoids): High-dose glucocorticoids are employed to modulate or prevent cytokine release syndrome induced by 145-2C11 administration.
  • Anti-TCR Antibodies: These may be used to monitor changes in TCR expression following treatment with 145-2C11.
  • TGF-? (Transforming Growth Factor Beta) Measurement: Levels of TGF-? are assessed to investigate the immunoregulatory or tolerogenic consequences of 145-2C11-induced T cell apoptosis.
  • Fc?R-related Antibodies or Ligands: Studies using Fc-modified or surrogate anti-CD3 antibodies (like 2C11-Novi) often involve comparisons with 145-2C11 to analyze Fc receptor interactions and downstream effects.
  • PLP139–151 Peptide: Used in experimental autoimmune encephalomyelitis (EAE) models to investigate the immunotherapeutic effects of anti-CD3 antibodies.

These antibodies and proteins are utilized in applications such as flow cytometry, immunohistochemistry, Western blotting, and in vivo functional studies. The precise combination depends on the experimental design, such as immune cell depletion, functional assays, cytokine profiling, or T cell activation studies.

Clone 145-2C11 is a widely used monoclonal antibody that targets the CD3? chain of the T cell receptor (TCR) complex in mice, and its scientific literature citations reveal several key findings regarding T cell activation, depletion, and immunomodulation.

Key findings:

  • Specificity and Function: 145-2C11 binds specifically to the 25-kDa ? chain of murine CD3, a crucial component of the TCR complex on thymocytes, mature T cells, and peripheral T cells. This binding is used routinely for identification and analysis of T cells by flow cytometry and other immunological assays.

  • T Cell Activation and Depletion: In vivo, administration of 145-2C11 potently activates T cells, leading to upregulation of activation markers such as CD69 and CD25, cytokine release (TNF, IFN?, IL-6), and T cell receptor (TCR) downregulation. 145-2C11 induces rapid, substantial TCR internalization, with recovery of surface expression occurring within five days after treatment. It also mediates T cell depletion via activation-induced cell death (AICD) and is more effective in this respect than Fc-modified variants.

  • Cytokine Release and Tolerogenic Effects: 145-2C11 administration results in robust cytokine secretion and can induce a tolerogenic (immune tolerance-promoting) response characterized by increased TGF-? levels. This property is linked to phagocyte-mediated clearance of apoptotic T cells, which is a source of tolerogenic cytokines.

  • Applications: The antibody is extensively used for:

    • Flow cytometry to detect murine T cells.
    • In vitro and in vivo T cell activation and proliferation assays.
    • T cell depletion in mouse models, both by apoptosis induction (especially in immature thymocytes) and by triggering activation-induced death.
    • Blocking CTL-mediated cytolysis and other functional studies of T cells.
    • Additional uses include immunoprecipitation, immunohistochemistry (when not formalin-fixed), and Western blotting.
  • Blocking Activity: 145-2C11 can block binding of other anti-CD3 antibodies (e.g., 17A2) to CD3?, making it useful in competitive or blocking experiments.

  • Historical Note: The clone was first described following immunization of Armenian hamsters with murine cytolytic T cells and identified as specific for the murine T3 (CD3) complex.

Summary Table:

Finding/UseEvidence
Binds murine CD3?/TCR complex
Used for T cell identification
Potent T cell activation in vivo
Induces AICD, T cell depletion
Increases cytokine (e.g., TGF-?)
Useful in various immunoassays
Blocks anti-CD3 antibody binding

These core findings make 145-2C11 a gold-standard tool in murine immunology research, especially for studies involving T cell function, modulation, and depletion.

References & Citations

1.) Abdulreda, M.H. et al. (2019) Diabetologia. 63, 237–1250
2.) Skyberg, J. A. et al. (2020) Infection and Immunity. 88: 5. Journal Link
Act
B
CyTOF®
Depletion
FA
Flow Cytometry
ICC
IF
IHC FF
in vivo Protocol
PhenoCycler®
General Western Blot Protocol

Certificate of Analysis

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