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Human peripheral blood mononuclear cells (PBMCs) were stained with CD133/1 (AC133) antibodies as well es with CD34 and CD45 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. A pre-gate of CD45+
cells was used. 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 tandems.
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