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Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (left image) or stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/mL ionomycin for 6 hours, followed by an incubation with 1 µg/mL brefeldin A after 2 hours at 37 °C. Cells were fixed, permeabilized, and stained with Anti-IL-2 antibodies as well as with CD69 antibodies. Flow cytometry was performed using the MACSQuant®
Analyzer. Cell debris were excluded from the analysis based on scatter signals.
In order to compare the epitope specificity of REAfinity Clone REA689 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 REA689|