Immunofluorescence analysis of CD3e was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD3e (OKT3) Mouse Monoclonal Antibody (MA110175) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous and cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Commercially sensitive information|
|Storage buffer||PBS, pH 7.4|
|Contains||15mM sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Functional Assay (FN)||Assay Dependent|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||Assay Dependent|
* 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
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.
Peripheral immature CD2-/low T cell development from type 2 to type 1 cytokine production.
MA1-10175 was used in flow cytometry to identify and characterize immature CD2(-/low) T cells in peripheral blood.
|Loza MJ,Perussia B||Journal of immunology (Baltimore, Md. : 1950) (169:3061)||2002|