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Human peripheral blood cells after erythrocyte lysis were stained with CD66b 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. 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.
REAfinity clone is tested against different known clones, in a blocking experiment, to identify whether they recognize completely overlapping (++), partially overlapping (+), or completely different epitopes (-). In a blocking experiment, cells are incubated with an excess of purified unconjugated REAfinity clone followed by staining with conjugated form of the other known clones.
|Other clones||Overlap in epitope recognition with REA306|
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