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Purified anti-CD3ε Antibody

Pricing & Availability
Clone
W23186F (See other available formats)
Regulatory Status
RUO
Other Names
T-cell surface glycoprotein CD3 epsilon chain, T3
Isotype
Rat IgG2a, κ
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Product Citations
publications
A. W23186F_PURE_CD3e_IHC-P_1_102324
IHC staining with Purified anti-CD3ε (clone W23186F) on formalin-fixed paraffin-embedded human tonsil tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated without (panel A) or with (panel B) Purified anti-CD3ε (clone W23186F), followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Images were captured with a 40X objective and merged. Scale bar: 50 µm
  • A. W23186F_PURE_CD3e_IHC-P_1_102324
    IHC staining with Purified anti-CD3ε (clone W23186F) on formalin-fixed paraffin-embedded human tonsil tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated without (panel A) or with (panel B) Purified anti-CD3ε (clone W23186F), followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Images were captured with a 40X objective and merged. Scale bar: 50 µm
  • B. W23186F_PURE_CD3e_2_IHC-P_102324
    IHC staining with Purified anti-CD3ε (clone W23186F) on formalin-fixed paraffin-embedded human tonsil tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated with Purified rat anti-CD3ε (clone W23186F) (red) and Purified anti-human CD8a (green) (Cat. No. 372902), followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) and Alexa Fluor® 594 Goat anti-mouse IgG (Cat. No. 405326) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (blue). Scale bar: Scale bar: 50 µm
  • C. W23186F_PURE_CD3e_WB_1_102324
    Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-CD3ε (clone W23186F) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-Rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:10000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • D. W23186F_PURE_CD3e_WB_2_102324
    Whole cell extracts (15 µg total protein per lane) from mouse spleen were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-CD3ε (clone W23186F) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-Rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:10000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • E. W23186F_PURE_CD3e_ICFC_102324
    Human peripheral blood lymphocytes were fixed and permeabilized using Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803), intracellularly stained with either Purified anti-CD3ε (clone W23186F), or Purified anti-rat IgG2a, κ isotype control (clone RTK2758) (Cat. No. 400502), and co-stained with PE anti-human CD19 (clone HIB19) (Cat. No. 302254).
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649951 25 µg £76
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649952 100 µg £189
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Description

The CD3ε chain of the T-cell receptor (TCR)-CD3 complex is a type I membrane protein that plays a critical role in antigen recognition, TCR signal transduction and T-cell activation following binding to the peptide/MHC antigen complex. CD3ε is expressed by all mature T cells and forms a heteromeric complex with CD3 γ, δ and ζ chains, and either α/β or γ/δ chains of the TCR heterodimer. Upon activation, CD3 undergoes a conformational change, leading to exposure of the proline rich sequence in CD3ε and recruitment of Lck. CD3 is also important in T cell development, and deficiency of this gene has been associated with severe immunodeficiency. 

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant fragment of the intracellular domain of human CD3ε
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

IHC-P - Quality tested
WB, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 1.0 - 10.0 µg/mL is suggested. For western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

For use in immunohistochemistry on formalin-fixed paraffin-embedded tissue (IHC-P), it is recommended to perform antigen retrieval using Citrate Buffer, 10X (Cat. No. 420902) or Tris-EDTA pH 9.0 Antigen Retrieval Buffer (10X) (Cat. No. 422704).

For use in intracellular flow cytometry (ICFC), fixation/permeabilization with Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) is recommended. Permeabilization with True-Phos™ Perm Buffer is not recommended.

This antibody (clone W23186F) is not recommended for use in immunocytochemistry (ICC) due to dim signal.

Antigen Details

Structure
CD3ε is a 207 amino acid protein with a predicted molecular weight of 23 kD.
Distribution

Plasma membrane

Function
Co-receptor
Interaction
CD3ε forms a heteromeric complex with CD3 γ, δ.and ζ chains, and either α/β or γ/δ chains of the TCR heterodimer.
Ligand/Receptor
CD3ε, as part of the TCR-CD3 complex, recognizes fragments of antigens bound to major histocompatibility complex molecules
Cell Type
NKT cells, T cells, Thymocytes, Tregs
Biology Area
Immunology
Molecular Family
CD Molecules, TCRs
Antigen References
  1. Schlossman S, et al. 1995. Leucocyte Typing V Oxford University Press New York.
  2. Knapp W. 1989. Leucocyte Typing IV. Oxford University Press New York.
  3. Barclay N, et al. 1997. The Leucocyte Antigen Facts Book. Academic Press Inc. San Diego.
  4. Sedelies KA, et al. 2004. J Biol Chem. 279:26581-7. 
  5. Rivollier A, et al. 2004. Blood. 104:4029-37. 
  6. Scharschmidt E, et al. 2004. Mol. Cell Biol. 24:3860-73. 
  7. Smeltz RB. 2007. J Immunol. 178:4786-92.
Gene ID
12501 View all products for this Gene ID
UniProt
View information about CD3epsilon on UniProt.org
Go To Top Version: 1    Revision Date: 10/23/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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