BD HorizonTM BV421 Rat Anti-Mouse TNF

Description

Two color flow cytometric analysis of TNF expression by stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma 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 Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse TNF antibody Cat. No. 563387; Right Panel by using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing correlated expression of TNF or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Two color flow cytometric analysis of TNF expression by stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma 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 Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse TNF antibody Cat. No. 563387; Right Panel by using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing correlated expression of TNF or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Two color flow cytometric analysis of TNF expression by stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma 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 Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse TNF antibody Cat. No. 563387; Right Panel by using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing correlated expression of TNF or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Show More Two color flow cytometric analysis of TNF expression by stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma 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 Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse TNF antibody Cat. No. 563387; Right Panel by using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing correlated expression of TNF or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Show More Two color flow cytometric analysis of TNF expression by stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma 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 Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse TNF antibody Cat. No. 563387; Right Panel by using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing correlated expression of TNF or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Two color flow cytometric analysis of TNF expression by stimulated mouse splenocytes. Mouse splenic leucocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate PMA; Sigma P-8139; 50 ng/ml and Ionomycin Sigma 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 Rat Anti-Mouse CD4 antibody Cat. No. 553051/561091 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse TNF antibody Cat. No. 563387; Right Panel by using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing correlated expression of TNF or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System.