BD HorizonTM BB700 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, washed with stain buffer, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 , and then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Rat Anti-Mouse TNF antibody Cat. No. 566510/566511; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing the 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 X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific. 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, washed with stain buffer, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 , and then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Rat Anti-Mouse TNF antibody Cat. No. 566510/566511; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing the 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 X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific. 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, washed with stain buffer, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 , and then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Rat Anti-Mouse TNF antibody Cat. No. 566510/566511; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing the 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 X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific. 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, washed with stain buffer, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 , and then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Rat Anti-Mouse TNF antibody Cat. No. 566510/566511; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing the 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 X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific. 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, washed with stain buffer, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 , and then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Rat Anti-Mouse TNF antibody Cat. No. 566510/566511; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing the 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 X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific. 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, washed with stain buffer, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were fixed and permeabilized with BD Cytofix/Cytoperm Fixation and Permeabilization Solution Cat. No. 554722 , and then washed and stained in BD Perm/Wash Buffer Cat. No. 554723 with FITC Rat Anti-Mouse CD4 antibody Cat. No. 553047/553046/561835 and either BD Horizon BB700 Rat IgG1, kappa Isotype Control Cat. No. 566420; Left Plot or BD Horizon BB700 Rat Anti-Mouse TNF antibody Cat. No. 566510/566511; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Flow cytometric dot plots showing the 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 X-20 Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.