BD HorizonTM RY586 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 fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . After washing the cells were permeabilized with BD Perm/Wash Buffer Cat. No. 554723 then stained in this buffer with APC Rat Anti-Mouse CD4 antibody Cat. No. 561091/553051 and with either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse TNF antibody Cat. No. 568547/568548; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD4 was 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 and FlowJo software. 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 fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . After washing the cells were permeabilized with BD Perm/Wash Buffer Cat. No. 554723 then stained in this buffer with APC Rat Anti-Mouse CD4 antibody Cat. No. 561091/553051 and with either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse TNF antibody Cat. No. 568547/568548; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD4 was 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 and FlowJo software. 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 fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . After washing the cells were permeabilized with BD Perm/Wash Buffer Cat. No. 554723 then stained in this buffer with APC Rat Anti-Mouse CD4 antibody Cat. No. 561091/553051 and with either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse TNF antibody Cat. No. 568547/568548; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD4 was 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 and FlowJo software. 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 fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . After washing the cells were permeabilized with BD Perm/Wash Buffer Cat. No. 554723 then stained in this buffer with APC Rat Anti-Mouse CD4 antibody Cat. No. 561091/553051 and with either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse TNF antibody Cat. No. 568547/568548; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD4 was 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 and FlowJo software. 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 fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . After washing the cells were permeabilized with BD Perm/Wash Buffer Cat. No. 554723 then stained in this buffer with APC Rat Anti-Mouse CD4 antibody Cat. No. 561091/553051 and with either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse TNF antibody Cat. No. 568547/568548; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD4 was 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 and FlowJo software. 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 fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . After washing the cells were permeabilized with BD Perm/Wash Buffer Cat. No. 554723 then stained in this buffer with APC Rat Anti-Mouse CD4 antibody Cat. No. 561091/553051 and with either BD Horizon RY586 Rat IgG1, kappa Isotype Control Cat. No. 568822; Left Plot or BD Horizon RY586 Rat Anti-Mouse TNF antibody Cat. No. 568547/568548; Right Plot at 0.25 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. The bivariate pseudocolor density plot showing the correlated expression of TNF or Ig Isotype control staining versus CD4 was 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 and FlowJo software. Data shown on this Technical Data Sheet are not lot specific.