Anti-CD3 zeta (CD247) Antibody

T-cell surface glycoprotein CD3 zeta chain (UniProt: P20963; also known as T-cell receptor T3 zeta chain, CD247) is encoded by the CD247 (also known as CD3Z, T3Z, TCRZ) gene (Gene ID: 919) in human. CD3 zeta is a single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to generate the mature form that contains a short extracellular domain (aa 22-30), a transmembrane domain (aa 31-51), and a cytoplasmic domain (aa 52-164). CD3 zeta is expressed in normal lymphoid tissues and in peripheral blood mononuclear cells (PBMC). It is a part of the TCR-CD3 complex that is present of the surface of T-cells and plays an essential role in adaptive immune response. When antigen presenting cells (APCs) activate T-cell receptor (TCR), TCR-mediated signals are transmitted across the cell membrane by various CD3 chains (CD3D, CD3E, CD3G and CD3Z). These chains contain immunoreceptor tyrosine-based activation motifs (ITAMs) in their cytoplasmic domain. The ITAM motifs CD3 zeta are localized to amino acids 61-89; 100-128; and 131-159. These motifs are reported to be phosphorylated by Src family protein tyrosine kinases Lck and Fyn, resulting in the activation of downstream signaling pathways. In CD3 zeta, phosphorylation of ITAMs by Lck is reported to be an initial step in the transduction of signaling cascades. It creates multiple docking sites for protein kinase ZAP70, which is also phosphorylated by Lck. Phosphorylation of ZAP70 converts it into a catalytically active form. It is reported that CD3 zeta phosphorylation can also be promoted by ZAP70 and TCR-induced zeta phosphorylation is shown to be defective in ZAP70-deficient T-cells.

Clone 6B10.2 is a mouse monoclonal antibody that detects CD3 zeta (CD247). It targets an epitope within 19 amino acids from the cytoplasmic domain.

KLH-conjugated linear peptide corresponding to 19 amino acids from the cytoplasmic domain of human CD3 zeta (CD247).

Quality Control Testing

Evaluated by Western Blotting in Jurkat cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected CD3 zeta (CD247) in Jurkat cell lysate.

Tested Applications

Flow Cytometry Analysis: A representative lot detected CD3 zeta (CD247) in Flow Cytometry applications (Machado Diaz, A.C., et. al. (2012). Arthritis. 2012:943156).

Immunoprecipitation Analysis: A representative lot immunoprecipitated CD3 zeta (CD247) in Immunoprecipitation applications (van Oers, N.S., et. al. (1995). J Exp Med. 182(5):1585-90).

Immunohistochemistry Applications: A representative lot detected CD3 zeta (CD247) in Immunohistochemistry applications (Balkhi, M.Y., et. al. (2018). iScience. 2:105-122).

Immunofluorescence Analysis: A representative lot detected CD3 zeta (CD247) in Immunofluorescence applications (Lee, J., et. al. (2020). NPJ Biofilms Microbiomes. 6(1):49).

Western Blotting Analysis: A representative lot detected CD3 zeta (CD247) in Western Blotting applications (Aki, D., et. al. (2018). Nat Immunol. 19(7):766-775; van Oers, N.S., et. al. (1995). J Exp Med. 182(5):1585-90).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

This mouse monoclonal Anti-CD3 zeta (CD247), clone 6B10.2, Cat. No. MABF3032 is tested for use in Western Blotting, Immunofluorescence, Immunohistochemistry, Immunoprecipitation and Flow Cytometry, for the detection of CD3 zeta (CD247).

Purified mouse monoclonal antibody IgG1 in PBS without azide.

Stable for 1 year at -10°C to -25°C from date of receipt. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.