BD HorizonTM PE-CF594 Rat Anti-Human IL-4

Description

Expression of IL-4 by stimulated human peripheral blood CD4+ T cells. An enriched preparation of peripheral blood CD4+ T cells 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 proteins for 3 days. Finally, the cells were harvested and stimulated for 5 h 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, and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Mouse Anti-Human CD4 Cat. No. 561841/555349/561840 and either BD Horizon PE-CF594 Rat IgG1 kappa Isotype Control Cat. No. 562309; Left Panel or BD Horizon PE-CF594 Mouse Anti-Human IL-4 antibody Cat. No. 565161; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 were derived from 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 CD4+ T cells. An enriched preparation of peripheral blood CD4+ T cells 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 proteins for 3 days. Finally, the cells were harvested and stimulated for 5 h 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, and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Mouse Anti-Human CD4 Cat. No. 561841/555349/561840 and either BD Horizon PE-CF594 Rat IgG1 kappa Isotype Control Cat. No. 562309; Left Panel or BD Horizon PE-CF594 Mouse Anti-Human IL-4 antibody Cat. No. 565161; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 were derived from 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 CD4+ T cells. An enriched preparation of peripheral blood CD4+ T cells 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 proteins for 3 days. Finally, the cells were harvested and stimulated for 5 h 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, and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Mouse Anti-Human CD4 Cat. No. 561841/555349/561840 and either BD Horizon PE-CF594 Rat IgG1 kappa Isotype Control Cat. No. 562309; Left Panel or BD Horizon PE-CF594 Mouse Anti-Human IL-4 antibody Cat. No. 565161; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 were derived from 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 CD4+ T cells. An enriched preparation of peripheral blood CD4+ T cells 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 proteins for 3 days. Finally, the cells were harvested and stimulated for 5 h 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, and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Mouse Anti-Human CD4 Cat. No. 561841/555349/561840 and either BD Horizon PE-CF594 Rat IgG1 kappa Isotype Control Cat. No. 562309; Left Panel or BD Horizon PE-CF594 Mouse Anti-Human IL-4 antibody Cat. No. 565161; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 were derived from 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 CD4+ T cells. An enriched preparation of peripheral blood CD4+ T cells 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 proteins for 3 days. Finally, the cells were harvested and stimulated for 5 h 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, and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Mouse Anti-Human CD4 Cat. No. 561841/555349/561840 and either BD Horizon PE-CF594 Rat IgG1 kappa Isotype Control Cat. No. 562309; Left Panel or BD Horizon PE-CF594 Mouse Anti-Human IL-4 antibody Cat. No. 565161; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 were derived from 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 CD4+ T cells. An enriched preparation of peripheral blood CD4+ T cells 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 proteins for 3 days. Finally, the cells were harvested and stimulated for 5 h 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, and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with APC Mouse Anti-Human CD4 Cat. No. 561841/555349/561840 and either BD Horizon PE-CF594 Rat IgG1 kappa Isotype Control Cat. No. 562309; Left Panel or BD Horizon PE-CF594 Mouse Anti-Human IL-4 antibody Cat. No. 565161; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric contour plots showing the correlated expression of IL-4 or Ig Isotype control staining versus CD4 were derived from 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.