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Human peripheral blood mononuclear cells (PBMCs) were stained with CD1c (BDCA-1) antibodies or with the corresponding isotype control antibodies (left images) as well as with CD19 and CD11c antibodies and analyzed by flow cytometry using the MACSQuant®
Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. For all other conjugates the FcR Blocking Reagent has been used to avoid Fc receptor–mediated antibody labeling. Apart from type-1 myeloid dendritic cells (MDC1s), CD1c (BDCA-1) is also expressed on a subset of B cells. For this reason, B cells were excluded from the analysis according to expression of CD19 and further analysis is restricted to CD19–
cells. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
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