Description: This 6B10.2 monoclonal antibody reacts with human CD247, which is also known as the invariant zeta chain. The CD3 complex is composed of three monomeric chains (gamma, delta, and epsilon) and the invariant zeta chain, which exists as a homodimer or as a heterodimer with the epsilon subunit. Together with the T cell antigen receptor (TCR), the CD3 gamma, delta, epsilon, and zeta chains comprise the TCR/CD3 complex. CD3 zeta is expressed on immature and mature thymocytes, as well as T lymphocytes and NK cells. Signal transduction during T cell activation is mediated by tyrosine phosphorylation of cytoplasmic immunoreceptor tyrosine-based activation motifs (ITAMs) present on the zeta chain.
The 6B10.2 monoclonal antibody recognizes a transmembrane epitope of the CD3 zeta chain. Therefore, flow cytometric analysis with this antibody requires intracellular staining. In addition, this antibody has been reported to recognize the zeta chain as a homodimer but not as a monomer.
Applications Reported: This 6B10.2 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This 6B10.2 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of normal human peripheral blood cells using the Intracellular Fixation and Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This antibody can be used at 5 µL (0.03 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.
Filtration: 0.2 µm post-manufacturing filtered.
CD3 complex is crucial in transducing antigen-recognition signals into the cytoplasm of T cells and in regulating the cell surface expression of the TCR complex. T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits CD3 gamma, CD3 delta, CD3 epsilon and CD3 zeta. These CD3 subunits are structurally related members of the immunoglobulins super family encoded by closely linked genes on human chromosome 11. The CD3 components have long cytoplasmic tails that associate with cytoplasmic signal transduction molecules. This association is mediated at least in part by a double tyrosine-based motif present in a single copy in the CD3 subunits. CD3 may play a role in TCR-induced growth arrest, cell survival and proliferation. The CD3 antigen is present on 68-82% of normal peripheral blood lymphocytes, 65-85% of thymocytes and Purkinje cells in the cerebellum. It is never expressed on B or NK cells. Decreased percentages of T lymphocytes may be observed in some autoimmune diseases.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD247; CD247 antigen, zeta subunit; CD3-ZETA; CD3H; CD3Q; CD3Z antigen, zeta polypeptide (TiT3 complex); CD3zeta chain; T-cell antigen receptor complex, zeta subunit of CD3; T-cell receptor T3 zeta chain; T-cell surface glycoprotein CD3 zeta chain; T3Z; TCR zeta chain; TCRZ
Gene Aliases: CD247; CD3-ZETA; CD3H; CD3Q; CD3Z; IMD25; T3Z; TCRZ
UniProt ID: (Human) P20963
Entrez Gene ID: (Human) 919