Key Product Details

Species Reactivity

Human

Applications

Multiplex Immunofluorescence, Immunohistochemistry, Western Blot, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1033713
View all CD3 epsilon Primary Antibodies »

Product Specifications

Immunogen

Human embryonic kidney cell HEK293-derived human CD3 epsilon
Gln22-Asp126
Accession # P07766

Specificity

Detects human CD3 epsilon in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human CD3 epsilon Antibody

Detection of CD3ɛ in Human T-Cell Lymphoma via seqIF™ Staining on COMET™​

CD3ɛ was detected in immersion fixed paraffin-embedded sections of human T-Cell Lymphoma using Mouse Anti-Human CD3ɛ Monoclonal Antibody (Catalog # MAB10670) at 18ug/mL at 37 ° Celsius for 2 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane of T-cells. Protocol available in COMET™ Panel Builder.​​​

Detection of Human CD3 epsilon by Western Blot.

Western blot shows lysates of MOLT‑4 human acute lymphoblastic leukemia cell line and Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human CD3 epsilon Monoclonal Antibody (Catalog # MAB10670) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for CD3 epsilon at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

CD3 epsilon in Human Thymus.

CD3 epsilon was detected in immersion fixed paraffin-embedded sections of human thymus using Mouse Anti-Human CD3 epsilon Monoclonal Antibody (Catalog # MAB10670) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface in lymphocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human CD3 epsilon by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and MOLT‑4 human acute lymphoblastic leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for CD3 epsilon at approximately 30 kDa (as indicated) using 25 µg/mL of Mouse Anti-Human CD3 epsilon Monoclonal Antibody (Catalog # MAB10670). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.

Applications for Human CD3 epsilon Antibody

Application
Recommended Usage

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffinembedded sections of human thymus

Multiplex Immunofluorescence

18 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human T-cell Lymphoma

Simple Western

25 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and MOLT‑4 human acute lymphoblastic leukemia cell line

Western Blot

2 µg/mL
Sample: MOLT‑4 human acute lymphoblastic leukemia cell line and Jurkat human acute T cell leukemia cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD3 epsilon

CD3 epsilon (T-Cell Surface Glycoprotein CD3 Epsilon) is a type I transmembrane protein that belongs to the Ig (Immunoglobulin) superfamily. Human CD3 epsilon consists of 104 amino acid (aa) extracellular domain, 26 aa transmembrane domain and 55 aa cytoplasmic domain (1). With the extracellular domain, human CD3 epsilon shares 44.2% and 49% homology with mouse and rat CD3 epsilon respectively. There are four CD3 proteins: CD3 delta, CD3 epsilon, CD3 gamma, and CD3 zeta. All CD3 proteins contain ITAM (Immunoreceptor Tyrosine-based Activation Motifs) in the cytoplasmic tail, which becomes phosphorylated by Src family protein tyrosine kinases LCK and FYN upon TCR engagement (2). CD3 proteins form heterodimers of CD3 delta /CD3 epsilon and CD3 gamma /CD3 epsilon, which bind to TCR and form trimeric TCR alpha /CD3 epsilon /CD3 gamma and TCR beta /CD3 gamma /CD3 epsilon. The resulting heterohexamer further associates with CD3 zeta homodimer and forms TCR/CD3 signaling complex (3, 4). Similar complex is also formed with TCR gamma and TCR delta (5). CD3 epsilon plays essential role in adaptive immune response. Mutations in CD3 epsilon lead to SCID (severe combined immunodeficiency) (6).

References

  1. Gold, D.P. et al. (1986) Nature 321:431.
  2. Barber, E.K. et al. (1989) Proc. Nat. Acad. Sci. U.S.A. 86:3277.
  3. Manolios, N. et al. (1991) EMBO J 10:1643.
  4. Dietrich, J. et al. (1999) Eur J Immunol 29:1719.
  5. Pan, Q. et al. (2006) Mol Immunol 43:1741.
  6. Soudais, C. et al. (1993) Nat. Genet. 3:77.

Alternate Names

CD3e

Entrez Gene IDs

916 (Human); 12501 (Mouse); 102133065 (Cynomolgus Monkey)

Gene Symbol

CD3E

UniProt

P07766

Additional CD3 epsilon Products

Product Documents for Human CD3 epsilon Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CD3 epsilon Antibody

For research use only

Citations for Human CD3 epsilon Antibody

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Protocols

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