BD HorizonTM BV786 Rat Anti-Human IL-4

Description

Expression of IL-4 by 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 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and human 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 and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Mouse Anti-Human CD4 Cat. No. 555346/561005/561842 and either BD Horizon BV786 Rat IgG1 kappa Isotype Control Cat. No. 563847; Left Panel or BD Horizon BV786 Mouse Anti-Human IL-4 antibody Cat. No. 564113; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-4 or Ig Isotype control staining versus CD4 for gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD LSR II Flow Cytometer System. Expression of IL-4 by 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 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and human 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 and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Mouse Anti-Human CD4 Cat. No. 555346/561005/561842 and either BD Horizon BV786 Rat IgG1 kappa Isotype Control Cat. No. 563847; Left Panel or BD Horizon BV786 Mouse Anti-Human IL-4 antibody Cat. No. 564113; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-4 or Ig Isotype control staining versus CD4 for gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD LSR II Flow Cytometer System. Show More Expression of IL-4 by 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 for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and human 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 and washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 . The cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Mouse Anti-Human CD4 Cat. No. 555346/561005/561842 and either BD Horizon BV786 Rat IgG1 kappa Isotype Control Cat. No. 563847; Left Panel or BD Horizon BV786 Mouse Anti-Human IL-4 antibody Cat. No. 564113; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-4 or Ig Isotype control staining versus CD4 for gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes. Flow cytometric analysis was performed using a BD LSR II Flow Cytometer System.