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Human peripheral blood mononuclear cells (PBMCs) were labeled with CD4-Biotin or unlabeled (left image) and fluorescently stained with Anti-Biotin antibodies as well as with CD3 antibodies. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. 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.