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Multicolor flow cytometric analysis of CD3 expression on viable Mouse splenic leukocytes. BALB/c Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) [Cat. No. 553142] and BD Pharmingen™ Purified Armenian Hamster Anti-Mouse FcγRIV (CD16-2) [Cat. No. 567208) antibodies. The leukocytes were then stained with APC Rat Anti-Mouse CD19 antibody (Cat. No. 550992) and with either BD Horizon™ RY610 Rat IgG2b, κ Isotype Control (Cat. No. 571153; Left Plot) or BD Horizon™ RY610 Rat Anti-Mouse CD3 antibody (Cat. No. 571224/571289; Right Plot) at 0.125 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD3 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ RY610 Rat Anti-Mouse CD3
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
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Companion Products
The 17A2 monoclonal antibody specifically binds to the T-cell receptor-associated CD3 complex that is expressed on many thymocytes and mature T lymphocytes. Plate-bound 17A2 antibody has been reported to induce IL-2 production by cultured T cells in the absence of accessory cells. The binding of the 17A2 antibody to T cells can be blocked by the anti-CD3e mAb 145-2C11 (Cat. No. 557306/553058/550275). This suggests that the 17A2 antibody recognizes an epitope of the CD3 epsilon chain. In vivo treatment with 17A2 antibody has been reported to partially deplete T lymphocytes and temporarily down-modulates CD3 expression on T cells.
Development References (4)
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Breitbach M, Kimura K, Luis TC, et al. In Vivo Labeling by CD73 Marks Multipotent Stromal Cells and Highlights Endothelial Heterogeneity in the Bone Marrow Niche.. Cell Stem Cell. 2018; 22(2):262-276.e7. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Miescher GC, Schreyer M, MacDonald HR. Production and characterization of a rat monoclonal antibody against the murine CD3 molecular complex. Immunol Lett. 1989; 23(2):113-118. (Immunogen: Cytotoxicity, Flow cytometry, Functional assay, Immunohistochemistry, Immunoprecipitation, Stimulation). View Reference
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Mysliwietz J, Thierfelder S. Antilymphocytic antibodies and marrow transplantation. XII. Suppression of graft-versus-host disease by T-cell-modulating and depleting antimouse CD3 antibody is most effective when preinjected in the marrow recipient. Blood. 1992; 80(10):2661-2667. (Clone-specific: Depletion, Flow cytometry, Functional assay, In vivo exacerbation). View Reference
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Wu L, Antica M, Johnson GR, Scollay R, Shortman K. Developmental potential of the earliest precursor cells from the adult mouse thymus. J Exp Med. 1991; 174(6):1617-1627. (Clone-specific: Cell separation, Depletion). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.