Staining of BALB/c splenocytes with Anti-Human/Mouse CD45R (B220) FITC (Product # 11-0452) and 0.125 µg of Rat IgG2b K Isotype Control PerCP-eFluor® 710 (Product # 46-4031) (left) or 0.125 µg of Anti-Mouse CD3 PerCP-eFluor® 710 (right). Cells in the lymphocyte gate were used for analysis.
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rat / IgG2b, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||0.25 µg/test|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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.
Applications Tested: This 17A2 antibody has been tested by flow cytometric analysis of mouse spleen cells. This can be used at less than or equal to 0.25 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.
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.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL cell sample + 100 µL 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
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.
||Murine sclerodermatous graft-versus-host disease, a model for human scleroderma: cutaneous cytokines, chemokines, and immune cell activation.||Zhang Y,McCormick LL,Desai SR,Wu C,Gilliam AC||Journal of immunology (Baltimore, Md. : 1950) (168:3088)||2002|
|Not Applicable||Not Cited||Murine sclerodermatous graft-versus-host disease, a model for human scleroderma: cutaneous cytokines, chemokines, and immune cell activation.||Zhang Y,McCormick LL,Desai SR,Wu C,Gilliam AC||Journal of immunology (Baltimore, Md. : 1950) (168:3088)||2002|