BD HorizonTM RY586 Rat Anti-Mouse IFN-gamma

Description

Two-color flow cytometric analysis of IFN-gamma expression in stimulated Mouse splenocytes. BALB/c Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma I-0634; 1 mug/ml in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse IFN-gamma antibody Cat. No. 568549/568550; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-gamma or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of IFN-gamma expression in stimulated Mouse splenocytes. BALB/c Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma I-0634; 1 mug/ml in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse IFN-gamma antibody Cat. No. 568549/568550; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-gamma or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of IFN-gamma expression in stimulated Mouse splenocytes. BALB/c Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma I-0634; 1 mug/ml in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse IFN-gamma antibody Cat. No. 568549/568550; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-gamma or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Show More Two-color flow cytometric analysis of IFN-gamma expression in stimulated Mouse splenocytes. BALB/c Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma I-0634; 1 mug/ml in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse IFN-gamma antibody Cat. No. 568549/568550; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-gamma or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Show More Two-color flow cytometric analysis of IFN-gamma expression in stimulated Mouse splenocytes. BALB/c Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma I-0634; 1 mug/ml in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse IFN-gamma antibody Cat. No. 568549/568550; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-gamma or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of IFN-gamma expression in stimulated Mouse splenocytes. BALB/c Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma I-0634; 1 mug/ml in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse IFN-gamma antibody Cat. No. 568549/568550; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-gamma or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific.