Human PBMCs stained with: CD45 Pacific Orange™ (MHCD4530TR), CD4 PerCP-Cy®5.5 (A15858), CD20 APC, CD14 APC-Cy®7 (A15453), CD19 Pacific Green™ (C11210), CD3 Alexa Fluor® 700 (CD0329), HLA-DR PE-Cy®7 (A18558), CD8 Pacific Blue™ (MHCD0828), CD33 FITC (A16185), and CD11c PE (A18674) using the Attune® NxT Acoustic Focusing Cytometer with 405 nm excitation and 440/50 emission filter (Pacific Blue™), 512/25 emission filter (Pacific Green™), and 603/48 emission filter (Pacific Orange™); 488 nm excitation and 530/30 emission filter (FITC) and 695/50 emission filter (PerCP-Cy®5.5); 561 nm excitation and 585/16 emission filter (PE) and 780/60 emission filter (PE-Cy®7); 637 nm excitation and 660/20 emission filter (APC), 720/30 emission filter (Alexa Fluor®700), and 780/60 emission filter (APC-Cy®7). Within the CD3 negative CD45 positive gate, B cells can be identified based on expression of CD19 and CD20. Conventional dendritic cells in peripheral blood are generally negative for T and B cell lineage markers and co-express the integrin CD11c and HLA-DR. Values noted are a percent of the parent CD19 negative CD20 negative gate (top value, no parenthesis) or percent of FSC/SSC lymphocyte gate (bottom value, in parentheses).
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Conjugate||Alexa Fluor® 700|
|Storage buffer||PBS with BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||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.
The Alexa Fluor® 700 dye conjugate provides an excellent spectral match to the common long-wavelength excitation sources, with a high extinction coefficient.
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.
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