BD HorizonTM BB700 Rat Anti-Human IL-4

Description

Flow cytometric analysis of IL-4 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml for plate coating and soluble Purified NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 2 mug/ml antibodies, recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and human IL-4 25 ng/ml final concentration; Cat. No. 554605 proteins for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and IL-4 for 3 days. Finally, the cells were harvested and stimulated for 5 hr with Phorbol 12-Myristate 13-Acetate Sigma, Cat. No. P-8139; 50 ng/ml and Ionomycin Sigma, Cat. No. I-0634; 1 mug/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 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BUV395 Mouse Anti-Human CD4 antibody Cat. No. 564724 and with either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Mouse Anti-Human IL-4 antibody Cat. No. 567744/567745; Right Plot using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Flow Cytometer System and FlowJo software. Flow cytometric analysis of IL-4 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml for plate coating and soluble Purified NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 2 mug/ml antibodies, recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and human IL-4 25 ng/ml final concentration; Cat. No. 554605 proteins for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and IL-4 for 3 days. Finally, the cells were harvested and stimulated for 5 hr with Phorbol 12-Myristate 13-Acetate Sigma, Cat. No. P-8139; 50 ng/ml and Ionomycin Sigma, Cat. No. I-0634; 1 mug/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 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BUV395 Mouse Anti-Human CD4 antibody Cat. No. 564724 and with either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Mouse Anti-Human IL-4 antibody Cat. No. 567744/567745; Right Plot using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Flow Cytometer System and FlowJo software. Flow cytometric analysis of IL-4 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml for plate coating and soluble Purified NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 2 mug/ml antibodies, recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and human IL-4 25 ng/ml final concentration; Cat. No. 554605 proteins for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and IL-4 for 3 days. Finally, the cells were harvested and stimulated for 5 hr with Phorbol 12-Myristate 13-Acetate Sigma, Cat. No. P-8139; 50 ng/ml and Ionomycin Sigma, Cat. No. I-0634; 1 mug/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 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BUV395 Mouse Anti-Human CD4 antibody Cat. No. 564724 and with either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Mouse Anti-Human IL-4 antibody Cat. No. 567744/567745; Right Plot using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Flow Cytometer System and FlowJo software. Show More Flow cytometric analysis of IL-4 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml for plate coating and soluble Purified NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 2 mug/ml antibodies, recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and human IL-4 25 ng/ml final concentration; Cat. No. 554605 proteins for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and IL-4 for 3 days. Finally, the cells were harvested and stimulated for 5 hr with Phorbol 12-Myristate 13-Acetate Sigma, Cat. No. P-8139; 50 ng/ml and Ionomycin Sigma, Cat. No. I-0634; 1 mug/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 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BUV395 Mouse Anti-Human CD4 antibody Cat. No. 564724 and with either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Mouse Anti-Human IL-4 antibody Cat. No. 567744/567745; Right Plot using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Flow Cytometer System and FlowJo software. Show More Flow cytometric analysis of IL-4 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml for plate coating and soluble Purified NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 2 mug/ml antibodies, recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and human IL-4 25 ng/ml final concentration; Cat. No. 554605 proteins for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and IL-4 for 3 days. Finally, the cells were harvested and stimulated for 5 hr with Phorbol 12-Myristate 13-Acetate Sigma, Cat. No. P-8139; 50 ng/ml and Ionomycin Sigma, Cat. No. I-0634; 1 mug/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 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BUV395 Mouse Anti-Human CD4 antibody Cat. No. 564724 and with either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Mouse Anti-Human IL-4 antibody Cat. No. 567744/567745; Right Plot using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Flow Cytometer System and FlowJo software. Flow cytometric analysis of IL-4 expression in stimulated human peripheral blood mononuclear cells. Human PBMC were stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml for plate coating and soluble Purified NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 2 mug/ml antibodies, recombinant human IL-2 10 ng/ml final concentration; Cat. No. 554603 and human IL-4 25 ng/ml final concentration; Cat. No. 554605 proteins for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and IL-4 for 3 days. Finally, the cells were harvested and stimulated for 5 hr with Phorbol 12-Myristate 13-Acetate Sigma, Cat. No. P-8139; 50 ng/ml and Ionomycin Sigma, Cat. No. I-0634; 1 mug/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 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BUV395 Mouse Anti-Human CD4 antibody Cat. No. 564724 and with either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Mouse Anti-Human IL-4 antibody Cat. No. 567744/567745; Right Plot using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Flow Cytometer System and FlowJo software.