BD PharmingenTM PE-Cy7 Rat Anti-Human IL-10

Description

Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody Cat. No. 555343 . The cells were cultured 5 d with plate-bound NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml, coated overnight at 4C and soluble NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 1 mug/ml antibodies plus recombinant Human IL-2 Cat. No. 554603; 20 ng/ml and IL-4 Cat. No. 554605; 40 ng/ml proteins. The cells were restimulated 5 h with Phorbol 12-Myristate 13-Acetate PMA; Sigma P8139; 50 ng/ml and Ionomycin Sigma I9657; 1 mug/ml and BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 . The cells were harvested, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BV421 Mouse Anti- Human CD4 antibody Cat. No. 565997/566907 and with either PE-Cy7 Rat IgG2a, kappa Isotype Control Cat. No. 552784; Left Plot or PE-Cy7 Mouse Anti-Human IL-10 antibody Cat. No. 567407/567408; Right Plot . The bivariate pseudocolor density plot showing the correlated expression of IL-10 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody Cat. No. 555343 . The cells were cultured 5 d with plate-bound NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml, coated overnight at 4C and soluble NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 1 mug/ml antibodies plus recombinant Human IL-2 Cat. No. 554603; 20 ng/ml and IL-4 Cat. No. 554605; 40 ng/ml proteins. The cells were restimulated 5 h with Phorbol 12-Myristate 13-Acetate PMA; Sigma P8139; 50 ng/ml and Ionomycin Sigma I9657; 1 mug/ml and BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 . The cells were harvested, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BV421 Mouse Anti- Human CD4 antibody Cat. No. 565997/566907 and with either PE-Cy7 Rat IgG2a, kappa Isotype Control Cat. No. 552784; Left Plot or PE-Cy7 Mouse Anti-Human IL-10 antibody Cat. No. 567407/567408; Right Plot . The bivariate pseudocolor density plot showing the correlated expression of IL-10 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody Cat. No. 555343 . The cells were cultured 5 d with plate-bound NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml, coated overnight at 4C and soluble NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 1 mug/ml antibodies plus recombinant Human IL-2 Cat. No. 554603; 20 ng/ml and IL-4 Cat. No. 554605; 40 ng/ml proteins. The cells were restimulated 5 h with Phorbol 12-Myristate 13-Acetate PMA; Sigma P8139; 50 ng/ml and Ionomycin Sigma I9657; 1 mug/ml and BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 . The cells were harvested, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BV421 Mouse Anti- Human CD4 antibody Cat. No. 565997/566907 and with either PE-Cy7 Rat IgG2a, kappa Isotype Control Cat. No. 552784; Left Plot or PE-Cy7 Mouse Anti-Human IL-10 antibody Cat. No. 567407/567408; Right Plot . The bivariate pseudocolor density plot showing the correlated expression of IL-10 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Show More Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody Cat. No. 555343 . The cells were cultured 5 d with plate-bound NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml, coated overnight at 4C and soluble NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 1 mug/ml antibodies plus recombinant Human IL-2 Cat. No. 554603; 20 ng/ml and IL-4 Cat. No. 554605; 40 ng/ml proteins. The cells were restimulated 5 h with Phorbol 12-Myristate 13-Acetate PMA; Sigma P8139; 50 ng/ml and Ionomycin Sigma I9657; 1 mug/ml and BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 . The cells were harvested, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BV421 Mouse Anti- Human CD4 antibody Cat. No. 565997/566907 and with either PE-Cy7 Rat IgG2a, kappa Isotype Control Cat. No. 552784; Left Plot or PE-Cy7 Mouse Anti-Human IL-10 antibody Cat. No. 567407/567408; Right Plot . The bivariate pseudocolor density plot showing the correlated expression of IL-10 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Show More Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody Cat. No. 555343 . The cells were cultured 5 d with plate-bound NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml, coated overnight at 4C and soluble NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 1 mug/ml antibodies plus recombinant Human IL-2 Cat. No. 554603; 20 ng/ml and IL-4 Cat. No. 554605; 40 ng/ml proteins. The cells were restimulated 5 h with Phorbol 12-Myristate 13-Acetate PMA; Sigma P8139; 50 ng/ml and Ionomycin Sigma I9657; 1 mug/ml and BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 . The cells were harvested, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BV421 Mouse Anti- Human CD4 antibody Cat. No. 565997/566907 and with either PE-Cy7 Rat IgG2a, kappa Isotype Control Cat. No. 552784; Left Plot or PE-Cy7 Mouse Anti-Human IL-10 antibody Cat. No. 567407/567408; Right Plot . The bivariate pseudocolor density plot showing the correlated expression of IL-10 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software. Two-color flow cytometric analysis of IL-10 expressed in stimulated human lymphocytes. CD4+ peripheral blood mononuclear cells were obtained by panning using plate-bound Purified NA/LE Mouse Anti-Human CD4 antibody Cat. No. 555343 . The cells were cultured 5 d with plate-bound NA/LE Mouse Anti-Human CD3 Cat. No. 555329; 10 mug/ml, coated overnight at 4C and soluble NA/LE Mouse Anti-Human CD28 Cat. No. 555725; 1 mug/ml antibodies plus recombinant Human IL-2 Cat. No. 554603; 20 ng/ml and IL-4 Cat. No. 554605; 40 ng/ml proteins. The cells were restimulated 5 h with Phorbol 12-Myristate 13-Acetate PMA; Sigma P8139; 50 ng/ml and Ionomycin Sigma I9657; 1 mug/ml and BD GolgiStop Protein Transport Inhibitor Cat. No. 554724 . The cells were harvested, fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , and permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with BD Horizon BV421 Mouse Anti- Human CD4 antibody Cat. No. 565997/566907 and with either PE-Cy7 Rat IgG2a, kappa Isotype Control Cat. No. 552784; Left Plot or PE-Cy7 Mouse Anti-Human IL-10 antibody Cat. No. 567407/567408; Right Plot . The bivariate pseudocolor density plot showing the correlated expression of IL-10 or Ig Isotype control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo software.