
563580 ·
BDB Bioscience · Cat: 563580
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Description
Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml 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 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 APC Mouse Anti-Human CD4 antibody Cat No. 561841/555349/561840 and either BD Horizon BV421 Rat IgG1 kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Mouse Anti-Human IL-13 antibody Cat. No. 563580; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 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 LSRFortessa Cell Analyzer System. Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml 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 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 APC Mouse Anti-Human CD4 antibody Cat No. 561841/555349/561840 and either BD Horizon BV421 Rat IgG1 kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Mouse Anti-Human IL-13 antibody Cat. No. 563580; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 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 LSRFortessa Cell Analyzer System. Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml 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 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 APC Mouse Anti-Human CD4 antibody Cat No. 561841/555349/561840 and either BD Horizon BV421 Rat IgG1 kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Mouse Anti-Human IL-13 antibody Cat. No. 563580; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 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 LSRFortessa Cell Analyzer System. Show More Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml 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 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 APC Mouse Anti-Human CD4 antibody Cat No. 561841/555349/561840 and either BD Horizon BV421 Rat IgG1 kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Mouse Anti-Human IL-13 antibody Cat. No. 563580; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 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 LSRFortessa Cell Analyzer System. Show More Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml 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 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 APC Mouse Anti-Human CD4 antibody Cat No. 561841/555349/561840 and either BD Horizon BV421 Rat IgG1 kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Mouse Anti-Human IL-13 antibody Cat. No. 563580; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 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 LSRFortessa Cell Analyzer System. Two-color flow cytometric analysis of IL-13 expression in stimulated human lymphocytes. Human peripheral blood mononuclear cells 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 and Recombinant Human IL-2 Cat. No. 554603; 10 ng/ml and Human IL-4 Cat. No. 554605; 25 ng/ml 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 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 APC Mouse Anti-Human CD4 antibody Cat No. 561841/555349/561840 and either BD Horizon BV421 Rat IgG1 kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Mouse Anti-Human IL-13 antibody Cat. No. 563580; Right Panel using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots show the correlated expression patterns of IL-13 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 LSRFortessa Cell Analyzer System.