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Human peripheral blood mononuclear cells (PBMCs) were labeled with CD303 (BDCA-2)-Biotin and fluorescently stained with Anti-Biotin antibodies as well as with CD304 (BDCA-4/Neuropilin-1) antibodies. 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. Cells were analyzed by flow cytometry using the MACSQuant®
Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
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