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Multicolor flow cytometric analysis of CD20 expression on mouse splenocytes. Mouse splenocytes from C57BL/6 mice were preincubated with Mouse BD Fc Block™ (Cat. No. 553141/553142). Cells were stained with BD Horizon™ BV421 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 562922) and with either BD Horizon™ R718 Rat IgG2a, κ Isotype Control (Cat No. 566941; Left Plot) or BD Horizon™ R718 Rat Anti-Mouse CD20 antibody (Cat No. 568258/568259; Right Plot) at 1.0 ug/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD45R/B220 signal was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) mouse splenocytes. 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™ R718 Rat Anti-Mouse CD20
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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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The GOT214A monoclonal antibody specifically binds to mouse CD20, also known as B-lymphocyte antigen and Bp35. CD20, a member of the MS4A gene family, is a 33-37 kDa protein comprised of four transmembrane spanning regions that present as a homo-oligomeric complexes on the cell surface. CD20 is expressed on most mature B cells but not on plasma cells. During development, CD20 is initially expressed after the induction of CD19 and is co-expressed with IgM during the pre-B to immature B cell transition in bone marrow. Expression of CD20 increases during maturation with almost all mature B cells expressing CD20 at some level. Unlike other B cell antigens, CD20 is not shed or internalized upon antibody binding. CD20 regulates B-cell activation and proliferation. Its ligation promotes transmembrane Ca2+ trafficking.
Development References (3)
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Tedder TF, Engel P. CD20: a regulator of cell-cycle progression of B lymphocytes.. Immunol Today. 1994; 15(9):450-4. (Biology). View Reference
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Uchida J, Hamaguchi Y, Oliver JA, et al. The innate mononuclear phagocyte network depletes B lymphocytes through Fc receptor-dependent mechanisms during anti-CD20 antibody immunotherapy.. J Exp Med. 2004; 199(12):1659-1669. (Biology: Flow cytometry). View Reference
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Uchida J, Lee Y, Hasegawa M, et al. Mouse CD20 expression and function.. Int Immunol. 2004; 16(1):119-29. (Biology). View Reference
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