Human CD3 epsilon Antibody

Catalog #: MAB100R 2 Reviews Datasheet

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB100R-100
MAB100R-SP
MAB100R-500
MAB100R-01M

Detection of Human CD3 epsilon by Western Blot.

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Human CD3 epsilon Antibody Summary

Species Reactivity

Human

Specificity

Recognizes the epsilon -chain of the CD3/T cell antigen receptor complex (McMichael, A.J. et al. (1987) Leucocyte Typing III: White Cell Differentiation Antigens, Oxford University Press, New York; Knapp, W. et al. (1989) Leucocyte Typing IV: White Cell Differentiation Antigens, Oxford University Press, New York; Schlossman, S. et al. (1995) Leucocyte Typing V: White Cell Differentiation Antigens, Oxford University Press, New York).

Source

Recombinant Monoclonal Mouse IgG₁ Clone # UCHT1R

Purification

Protein A or G purified from cell culture supernatant

Immunogen

Human thymocytes

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.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

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

Flow Cytometry

0.25 µg/10⁶ cells

See below

Immunocytochemistry

1-25 µg/mL

See below

T Cell Stimulation

This antibody can be used to activate T cells when immobilized at 1-10 μg/mL (100 μL/well).

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

Western Blot

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Detection of Human CD3 epsilon by Western Blot. Western blot shows lysates of MOLT‑4 human acute lymphoblastic leukemia cell line, Jurkat human acute T cell leukemia cell line, Raji human Burkitt's lymphoma cell line (negative control), and THP‑1 human acute monocytic leukemia cell line (negative control). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human CD3 epsilon Monoclonal Antibody (Catalog # MAB100R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for CD3 epsilon at approximately 21 kDa (as indicated). GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Flow Cytometry

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Detection of CD3 epsilon in Human Lymphocytes by Flow Cytometry. Human peripheral blood lymphocytes were stained with Mouse Anti-Human CD3e Recombinant Monoclonal Antibody (Catalog # MAB100, filled histogram) or isotype control antibody (MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (F0102B).

Immunocytochemistry

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CD3 epsilon in Human PBMCs. CD3e was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human CD3e Recombinant Monoclonal Antibody (Catalog # MAB100R) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; (NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

T Cell Stimulation

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T Cell Stimulation Induced by Human CD3 epsilon Antibody. Mouse Anti-Human CD3e Monoclonal Antibody (Catalog # MAB100R) induces stimulation in human T cells in a dose-dependent manner, as measured by Resazurin (AR002), when immobilized at 1-10 µg/mL (100 µL/well).

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

Background: CD3

CD3 epsilon is one of at least four invariant proteins that associate with the variable antigen recognition chains of the T cell receptor and function in signal transduction.

References

Beverly, P.C.L. and R.E. Callard (1981) Eur. J. Immunol. 11:329.

Entrez Gene IDs

916 (Human); 12501 (Mouse)

Alternate Names

CD_antigen: CD3e; CD3 antigen, delta subunit; CD3; CD3d antigen; CD3d antigen, delta polypeptide (TiT3 complex); CD3d molecule, delta (CD3-TCR complex); CD3-DELTA; CD3e antigen; CD3e antigen, epsilon polypeptide (TiT3 complex); CD3e molecule, epsilon (CD3-TCR complex); CD3e; CD3-epsilon; CD3g antigen; CD3g antigen, gamma polypeptide (TiT3 complex); CD3g molecule, epsilon (CD3-TCR complex); CD3g molecule, gamma (CD3-TCR complex); CD3G; CD3-GAMMA; FLJ17620; FLJ17664; FLJ18683; FLJ79544; FLJ94613; IMD18; MGC138597; T3DOKT3, delta chain; T3E; T-cell antigen receptor complex, epsilon subunit of T3; T-cell receptor T3 delta chain; T-cell surface antigen T3/Leu-4 epsilon chain; T-cell surface glycoprotein CD3 delta chain; T-cell surface glycoprotein CD3 epsilon chain; TCRE