BD HorizonTM R718 Rat Anti-Mouse IL-13

Description

Multiparameter flow cytometric analysis of IL-13 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. 567114/567115; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294/567110 antibodies with Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and Recombinant Mouse IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4 and 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 5.5 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553049/553048 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-13 antibody Cat. No. 569869/569945; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 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 X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Multiparameter flow cytometric analysis of IL-13 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. 567114/567115; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294/567110 antibodies with Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and Recombinant Mouse IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4 and 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 5.5 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553049/553048 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-13 antibody Cat. No. 569869/569945; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 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 X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Multiparameter flow cytometric analysis of IL-13 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. 567114/567115; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294/567110 antibodies with Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and Recombinant Mouse IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4 and 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 5.5 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553049/553048 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-13 antibody Cat. No. 569869/569945; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 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 X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Show More Multiparameter flow cytometric analysis of IL-13 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. 567114/567115; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294/567110 antibodies with Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and Recombinant Mouse IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4 and 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 5.5 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553049/553048 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-13 antibody Cat. No. 569869/569945; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 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 X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Show More Multiparameter flow cytometric analysis of IL-13 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. 567114/567115; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294/567110 antibodies with Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and Recombinant Mouse IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4 and 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 5.5 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553049/553048 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-13 antibody Cat. No. 569869/569945; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 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 X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Multiparameter flow cytometric analysis of IL-13 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. 567114/567115; 10 mug/ml for coating and soluble Purified NA/LE Hamster Anti-Mouse CD28 Cat. No. 553294/567110 antibodies with Recombinant Mouse IL-2 Cat. No. 550069; 10 ng/ml and Recombinant Mouse IL-4 Cat. No. 550067; 25 ng/ml proteins. The cells were subsequently cultured 3 days with Recombinant IL-2 and IL-4 and 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 5.5 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553049/553048 and with either BD Horizon R718 Rat IgG1, kappa Isotype Control Cat. No. 566948; Left Plot or BD Horizon R718 Rat Anti-Mouse IL-13 antibody Cat. No. 569869/569945; Right Plot at 0.125 mug/test using the BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of IL-13 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 X-20 Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific.