BD HorizonTM RB613 Rat Anti-Mouse TNF

Description

Two-color flow cytometric analysis of TNF expression in stimulated Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 with BD Cytofix Fixation Buffer Cat. No. 554655 . 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 and either BD Horizon RB613 Rat IgG1, kappa Isotype Control Cat. No. 571675; Left Plot or BD Horizon RB613 Rat Anti-Mouse TNF antibody Cat. No. 571272/571332; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density 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 leukocytes. Flow cytometric analysis was performed using a BD FACSymphony A5 SE Cell Analyzer and FlowJo Software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of TNF expression in stimulated Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 with BD Cytofix Fixation Buffer Cat. No. 554655 . 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 and either BD Horizon RB613 Rat IgG1, kappa Isotype Control Cat. No. 571675; Left Plot or BD Horizon RB613 Rat Anti-Mouse TNF antibody Cat. No. 571272/571332; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density 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 leukocytes. Flow cytometric analysis was performed using a BD FACSymphony A5 SE Cell Analyzer and FlowJo Software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of TNF expression in stimulated Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 with BD Cytofix Fixation Buffer Cat. No. 554655 . 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 and either BD Horizon RB613 Rat IgG1, kappa Isotype Control Cat. No. 571675; Left Plot or BD Horizon RB613 Rat Anti-Mouse TNF antibody Cat. No. 571272/571332; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density 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 leukocytes. Flow cytometric analysis was performed using a BD FACSymphony A5 SE Cell Analyzer and FlowJo Software. Data shown on this Technical Data Sheet are not lot specific. Show More Two-color flow cytometric analysis of TNF expression in stimulated Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 with BD Cytofix Fixation Buffer Cat. No. 554655 . 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 and either BD Horizon RB613 Rat IgG1, kappa Isotype Control Cat. No. 571675; Left Plot or BD Horizon RB613 Rat Anti-Mouse TNF antibody Cat. No. 571272/571332; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density 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 leukocytes. Flow cytometric analysis was performed using a BD FACSymphony A5 SE Cell Analyzer and FlowJo Software. Data shown on this Technical Data Sheet are not lot specific. Show More Two-color flow cytometric analysis of TNF expression in stimulated Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 with BD Cytofix Fixation Buffer Cat. No. 554655 . 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 and either BD Horizon RB613 Rat IgG1, kappa Isotype Control Cat. No. 571675; Left Plot or BD Horizon RB613 Rat Anti-Mouse TNF antibody Cat. No. 571272/571332; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density 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 leukocytes. Flow cytometric analysis was performed using a BD FACSymphony A5 SE Cell Analyzer and FlowJo Software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of TNF expression in stimulated Mouse splenic leukocytes. BALB/c Mouse splenocytes 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 with BD Cytofix Fixation Buffer Cat. No. 554655 . 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 and either BD Horizon RB613 Rat IgG1, kappa Isotype Control Cat. No. 571675; Left Plot or BD Horizon RB613 Rat Anti-Mouse TNF antibody Cat. No. 571272/571332; Right Plot at 0.5 mug/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density 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 leukocytes. Flow cytometric analysis was performed using a BD FACSymphony A5 SE Cell Analyzer and FlowJo Software. Data shown on this Technical Data Sheet are not lot specific.