BD HorizonTM RY586 Mouse Anti-Human TNF

Description

Flow cytometric analysis of TNF expression in stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells PBMC were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate Sigma P-8139; 50 ng/ml final concentration and Ionomycin Sigma I-0634; 1 mug/ml final concentration in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Mouse IgG1, kappa Isotype Control Cat No. 568097, Left Plot or with BD Horizon RY586 Mouse Anti-Human TNF antibody Cat No. 568461/568462, Right Plot . The pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD3 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 Flow Cytometer System and FlowJo software. Flow cytometric analysis of TNF expression in stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells PBMC were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate Sigma P-8139; 50 ng/ml final concentration and Ionomycin Sigma I-0634; 1 mug/ml final concentration in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Mouse IgG1, kappa Isotype Control Cat No. 568097, Left Plot or with BD Horizon RY586 Mouse Anti-Human TNF antibody Cat No. 568461/568462, Right Plot . The pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD3 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 Flow Cytometer System and FlowJo software. Flow cytometric analysis of TNF expression in stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells PBMC were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate Sigma P-8139; 50 ng/ml final concentration and Ionomycin Sigma I-0634; 1 mug/ml final concentration in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Mouse IgG1, kappa Isotype Control Cat No. 568097, Left Plot or with BD Horizon RY586 Mouse Anti-Human TNF antibody Cat No. 568461/568462, Right Plot . The pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD3 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 Flow Cytometer System and FlowJo software. Show More Flow cytometric analysis of TNF expression in stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells PBMC were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate Sigma P-8139; 50 ng/ml final concentration and Ionomycin Sigma I-0634; 1 mug/ml final concentration in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Mouse IgG1, kappa Isotype Control Cat No. 568097, Left Plot or with BD Horizon RY586 Mouse Anti-Human TNF antibody Cat No. 568461/568462, Right Plot . The pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD3 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 Flow Cytometer System and FlowJo software. Show More Flow cytometric analysis of TNF expression in stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells PBMC were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate Sigma P-8139; 50 ng/ml final concentration and Ionomycin Sigma I-0634; 1 mug/ml final concentration in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Mouse IgG1, kappa Isotype Control Cat No. 568097, Left Plot or with BD Horizon RY586 Mouse Anti-Human TNF antibody Cat No. 568461/568462, Right Plot . The pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD3 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 Flow Cytometer System and FlowJo software. Flow cytometric analysis of TNF expression in stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells PBMC were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate Sigma P-8139; 50 ng/ml final concentration and Ionomycin Sigma I-0634; 1 mug/ml final concentration in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The cells were then stained in BD Perm/Wash Buffer with BD Horizon BV421 Mouse Anti-Human CD3 antibody Cat. No. 562426 and with either BD Horizon RY586 Mouse IgG1, kappa Isotype Control Cat No. 568097, Left Plot or with BD Horizon RY586 Mouse Anti-Human TNF antibody Cat No. 568461/568462, Right Plot . The pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD3 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 Flow Cytometer System and FlowJo software.