BD PharmingenTM PE Mouse anti-Human IL-17F

Description

Flow cytometric analysis of IL-17F expression by resting and activated human peripheral blood CD4+ T cells and Th17 polarized cells. Human peripheral blood mononuclear cells were either unstimulated Left Panels or stimulated with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139 plus Ionomycin Sigma; I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 for 5 hours Middle Panels or were cultured in Th17 polarization conditions and restimulated with PMA and Ionomycin in the presence of BD GolgiStop for 5 hours Right Panels . Cells were then fixed and permeabilized using BD Cytofix/Cytoperm reagents Cat. No. 554714 followed by staining with PE Mouse anti-Human IL-17F Cat. No. 561197/561198 , PerCP-Cy5.5 Mouse anti-Human CD4 Cat. No. 341654 , and FITC Mouse Anti-Human IFN-gamma Cat. No. 554700 or Alexa Fluor 647 Mouse anti-Human IL-17A Cat. No. 560490 . Two-color flow cytometric dot plots showing the correlated expression patterns of IL-17F versus IL-17A or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Recommended Assay Procedure. Flow cytometric analysis of IL-17F expression by resting and activated human peripheral blood CD4+ T cells and Th17 polarized cells. Human peripheral blood mononuclear cells were either unstimulated Left Panels or stimulated with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139 plus Ionomycin Sigma; I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 for 5 hours Middle Panels or were cultured in Th17 polarization conditions and restimulated with PMA and Ionomycin in the presence of BD GolgiStop for 5 hours Right Panels . Cells were then fixed and permeabilized using BD Cytofix/Cytoperm reagents Cat. No. 554714 followed by staining with PE Mouse anti-Human IL-17F Cat. No. 561197/561198 , PerCP-Cy5.5 Mouse anti-Human CD4 Cat. No. 341654 , and FITC Mouse Anti-Human IFN-gamma Cat. No. 554700 or Alexa Fluor 647 Mouse anti-Human IL-17A Cat. No. 560490 . Two-color flow cytometric dot plots showing the correlated expression patterns of IL-17F versus IL-17A or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Recommended Assay Procedure. Show More Flow cytometric analysis of IL-17F expression by resting and activated human peripheral blood CD4+ T cells and Th17 polarized cells. Human peripheral blood mononuclear cells were either unstimulated Left Panels or stimulated with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139 plus Ionomycin Sigma; I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 for 5 hours Middle Panels or were cultured in Th17 polarization conditions and restimulated with PMA and Ionomycin in the presence of BD GolgiStop for 5 hours Right Panels . Cells were then fixed and permeabilized using BD Cytofix/Cytoperm reagents Cat. No. 554714 followed by staining with PE Mouse anti-Human IL-17F Cat. No. 561197/561198 , PerCP-Cy5.5 Mouse anti-Human CD4 Cat. No. 341654 , and FITC Mouse Anti-Human IFN-gamma Cat. No. 554700 or Alexa Fluor 647 Mouse anti-Human IL-17A Cat. No. 560490 . Two-color flow cytometric dot plots showing the correlated expression patterns of IL-17F versus IL-17A or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD LSR II Flow Cytometer System. Other compatible fixation and permeabilization treatments are listed in the Recommended Assay Procedure.