Human CD3 epsilon Antibody

Catalog #: MAB10670 Datasheet
Catalog # Availability Size / Price Qty
MAB10670-100
MAB10670-SP
Detection of CD3ɛ in Human T-Cell Lymphoma via seqIF™ Staining on COMET™​
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Human CD3 epsilon Antibody Summary

Species Reactivity
Human
Specificity
Detects human CD3 epsilon in direct ELISAs.
Source
Monoclonal Mouse IgG2B Clone # 1033713
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Human embryonic kidney cell HEK293-derived human CD3 epsilon
Gln22-Asp126
Accession # P07766
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
MOLT‑4 human acute lymphoblastic leukemia cell line and Jurkat human acute T cell leukemia cell line
Simple Western
25 µg/mL
Jurkat human acute T cell leukemia cell line and MOLT‑4 human acute lymphoblastic leukemia cell line
Immunohistochemistry
5-25 µg/mL
Immersion fixed paraffinembedded sections of human thymus

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Multiplex Immunofluorescence View Larger

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.​​​

Western Blot View Larger

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.

Immunohistochemistry View Larger

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.

Simple Western View Larger

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.

Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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.

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: CD3δ, 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 CD3δ/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 TCRδ (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.
Entrez Gene IDs
916 (Human); 12501 (Mouse); 102133065 (Cynomolgus Monkey)
Alternate Names
CD3 epsilon; CD3e antigen; CD3e antigen, epsilon polypeptide (TiT3 complex); CD3e molecule, epsilon (CD3-TCR complex); CD3e; CD3-epsilon; FLJ18683; T3E; T-cell antigen receptor complex, epsilon subunit of T3; T-cell surface antigen T3/Leu-4 epsilon chain; T-cell surface glycoprotein CD3 epsilon chain; TCRE

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