Description: The 17A2 monoclonal antibody reacts with the mouse CD3 complex. CD3 subunits gamma, delta and epsilon are required for proper assembly, trafficking and surface expression of the TCR complex. CD3 is expressed by thymocytes in a developmentally regulated manner and by all mature T cells. Binding of 17A2 to CD3 initiates the intracellular biochemical pathway resulting in cellular activation and proliferation.
Applications Reported: This 17A2 antibody has been reported for use in flow cytometric analysis and functional activity (in vitro simulation).
Applications Tested: This 17A2 antibody has been tested by flow cytometric analysis of mouse thymocytes and splenocytes. This may be used at less than or equal to 1 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
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 (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
The CD3 subunit complex which 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 and 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. The genes encoding the CD3 epsilon, gamma and delta polypeptides are located on chromosome 11. Defects in the CD3 gene are associated with CD3 immunodeficiency.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD247; CD247 antigen; CD3 antigen, zeta polypeptide; CD3-zeta/eta; CD3d; CD3e; CD3g; HCD3; T-cell receptor T3 delta chain; T-cell receptor T3 eta chain; T-cell receptor T3 gamma chain; T-cell receptor T3 zeta chain; T-cell surface antigen T3/Leu-4 epsilon chain; T-cell surface glycoprotein CD3 delta chain; T-cell surface glycoprotein CD3 epsilon chain; T-cell surface glycoprotein CD3 gamma chain; T-cell surface glycoprotein CD3 zeta chain
Gene Aliases: 4930549J05Rik; A430104F18Rik; AI504783; AW552088; Cd247; CD3; Cd3-eta; Cd3-zeta; Cd3d; Cd3e; CD3epsilon; Cd3g; Cd3h; Cd3z; Cd3zeta; Ctg-3; Ctg3; T3d; T3e; T3g; T3z; Tcrk; Tcrz