
568467 · 100 Tests
BDB Bioscience · Cat: 568467
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Description
Two-color flow cytometric analysis of IL-2 expression in activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Rat IgG2a, kappa Isotype Control Cat No. 568130, Left Plot or BD Horizon RY586 Rat Anti-Human IL-2 antibody Cat No. 568467/568468, Right Plot using BD Biosciences Intracellular Cytokine Staining Protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Two-color flow cytometric analysis of IL-2 expression in activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Rat IgG2a, kappa Isotype Control Cat No. 568130, Left Plot or BD Horizon RY586 Rat Anti-Human IL-2 antibody Cat No. 568467/568468, Right Plot using BD Biosciences Intracellular Cytokine Staining Protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Two-color flow cytometric analysis of IL-2 expression in activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Rat IgG2a, kappa Isotype Control Cat No. 568130, Left Plot or BD Horizon RY586 Rat Anti-Human IL-2 antibody Cat No. 568467/568468, Right Plot using BD Biosciences Intracellular Cytokine Staining Protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Show More Two-color flow cytometric analysis of IL-2 expression in activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Rat IgG2a, kappa Isotype Control Cat No. 568130, Left Plot or BD Horizon RY586 Rat Anti-Human IL-2 antibody Cat No. 568467/568468, Right Plot using BD Biosciences Intracellular Cytokine Staining Protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Show More Two-color flow cytometric analysis of IL-2 expression in activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Rat IgG2a, kappa Isotype Control Cat No. 568130, Left Plot or BD Horizon RY586 Rat Anti-Human IL-2 antibody Cat No. 568467/568468, Right Plot using BD Biosciences Intracellular Cytokine Staining Protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Two-color flow cytometric analysis of IL-2 expression in activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Rat IgG2a, kappa Isotype Control Cat No. 568130, Left Plot or BD Horizon RY586 Rat Anti-Human IL-2 antibody Cat No. 568467/568468, Right Plot using BD Biosciences Intracellular Cytokine Staining Protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software.