Flow cytometric comparison of different clones for IL-5. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-5 antibodies. As a control, IL-5 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for IL-5. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-5 antibodies. As a control, IL-5 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for IL-5. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-5 antibodies. As a control, IL-5 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for IL-5. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-5 antibodies. As a control, IL-5 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for IL-5. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL Brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-5 antibodies. As a control, IL-5 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.