Flow cytometric comparison of different clones for T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility^TM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. 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 T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility^TM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. 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 T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility^TM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. 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 T-bet. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility^TM Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the FoxP3 Staining Buffer Set followed by intracellular staining with T-bet antibodies. As a control, T-bet antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye-conjugated antibodies. 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.