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Human peripheral blood mononuclear cells (PBMCs) were stained with CD45RO antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD45RA antibodies. Flow cytometry was performed 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 REA611|
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