Human peripheral blood mononuclear cells were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate and Ionomycin in the presence of BD GolgiStop™ Protein Transport Inhibitor. The cells were permeabilized with BD Perm/Wash™ Buffer then stained with BD Horizon™ R718 Mouse Anti-Human IL-17A (clone N49-653, A) or BD Pharmingen™ Alexa Fluor™ 700 Mouse Anti-Human IL-17A (clone N49-653, B) and BD Pharmingen™ PE Mouse Anti-Human CD4 (clone RPA-T4) shown as two-color flow cytometric pseudocolor density plots derived from gated lymphocytes and were analyzed the same day on a BD LSRFortessa™ X-20 Cell Analyzer with FlowJo™ Software.

Comparison staining of CD4 on human lysed whole blood. A. Lysed whole blood stained with BD Horizon™ R718 Mouse Anti-Human CD4 (clone SK3, red) or BD Pharmingen™ Alexa Fluor™ 700 Mouse Anti-Human CD4 (clone SK3, grey) shown overlaid as fluorescence histograms derived from a lymphocyte gate. B. SSC vs. fluorescence plots of BD Horizon™ R718 Mouse Anti-Human CD4 (left) or BD Pharmingen™ Alexa Fluor™ 700 Mouse Anti-Human CD4 (right). All data were analyzed on a BD LSRFortessa™ X-20 Cell Analyzer with FlowJo™ Software

A. BD Horizon™ R718 FoxP3 is on par with the gold standard BD Pharmingen™ PE FoxP3. Human PBMCs were stained with BD Horizon™ R718 Mouse Anti-Human FoxP3 (clone 259D/C7). B. BD Phosflow™ Treated Human Control Cells (red) and BD Phosflow™ Untreated Human Control Cells (grey) were stained with BD Phosflow™ R718 Mouse Anti-Stat5 (pY694) (clone 47); treated and untreated control cell overlays shown here are derived from a CD4+ gate.