BD HorizonTM R718 Mouse Anti-Human IL-17A

Description

Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Show More Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Show More Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Show More Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate PMA; 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 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 Cat No. 564724 , FITC Mouse Anti-Human CD45RO Cat. No. 555492; Left Plot , BD Horizon BV421 Mouse Anti-Human IL-4 Cat. No. 564110; Middle Plot Plot , and PE Mouse Anti-Human IFN-gamma Cat. No. 559326; Right Plot Plot antibodies and with either BD Horizon R718 Rat IgG1 kappa Isotype Control Cat. No. 566928; Data Not Shown or BD Horizon R718 Mouse Anti-Human IL-17 antibody Cat. No. 567094/567247 using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-gamma were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software.