BD HorizonTM R718 Rat Anti-Mouse IL-4

Description

Two-color flow cytometric analysis of IL-4 expression in stimulated mouse T cells. Mouse splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 for 6 hours. The cells were washed and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were washed and then permeabilized and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-4 antibody Cat. No. 567083; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. A 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 intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of IL-4 expression in stimulated mouse T cells. Mouse splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 for 6 hours. The cells were washed and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were washed and then permeabilized and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-4 antibody Cat. No. 567083; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. A 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 intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of IL-4 expression in stimulated mouse T cells. Mouse splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 for 6 hours. The cells were washed and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were washed and then permeabilized and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-4 antibody Cat. No. 567083; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. A 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 intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa 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 IL-4 expression in stimulated mouse T cells. Mouse splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 for 6 hours. The cells were washed and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were washed and then permeabilized and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-4 antibody Cat. No. 567083; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. A 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 intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa 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 IL-4 expression in stimulated mouse T cells. Mouse splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 for 6 hours. The cells were washed and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were washed and then permeabilized and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-4 antibody Cat. No. 567083; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. A 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 intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of IL-4 expression in stimulated mouse T cells. Mouse splenic CD4+ T cells were cultured 2 days with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e Cat. No. 553057; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294; 2 mug/ml antibodies with Recombinant IL-2 Cat. No. 550069; 10 ng/ml and IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4. The cells were then stimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634 in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 for 6 hours. The cells were washed and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The fixed cells were washed and then permeabilized and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-4 antibody Cat. No. 567083; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. A 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 intact stimulated lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific.