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Human peripheral blood mononuclear cells (PBMCs) were stained with CD94 antibodies as well as with CD56 antibodies and analyzed by flow cytometry using the MACSQuant®
Analyzer. Lymphocytes were gated for the analysis. 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.
In order to compare the epitope specificity of REAfinity Clone REA113 with other known clones, a competition assay was performed.
Cells were incubated with an excess of purified unconjugated REAfinity Antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.
|Other clones||Overlap in epitope recognition with REA113|