BD HorizonTM RB705 Rat Anti-Mouse TNF

Description

Two-color flow cytometric analysis of TNF expression by stimulated Mouse splenic leukocytes. 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, washed with Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were washed and then permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with PE Rat Anti-Mouse CD4 antibody Cat. No. 553049 and with either no antibody BD Horizon RB705 Autofluorescence Control; Left Plot or BD Horizon RB705 Rat Anti-Mouse TNF antibody Cat. No. 570733/570734; 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 Autofluorescence control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were 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 splenic leukocytes. 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, washed with Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were washed and then permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with PE Rat Anti-Mouse CD4 antibody Cat. No. 553049 and with either no antibody BD Horizon RB705 Autofluorescence Control; Left Plot or BD Horizon RB705 Rat Anti-Mouse TNF antibody Cat. No. 570733/570734; 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 Autofluorescence control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were 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 splenic leukocytes. 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, washed with Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were washed and then permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with PE Rat Anti-Mouse CD4 antibody Cat. No. 553049 and with either no antibody BD Horizon RB705 Autofluorescence Control; Left Plot or BD Horizon RB705 Rat Anti-Mouse TNF antibody Cat. No. 570733/570734; 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 Autofluorescence control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were 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 splenic leukocytes. 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, washed with Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were washed and then permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with PE Rat Anti-Mouse CD4 antibody Cat. No. 553049 and with either no antibody BD Horizon RB705 Autofluorescence Control; Left Plot or BD Horizon RB705 Rat Anti-Mouse TNF antibody Cat. No. 570733/570734; 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 Autofluorescence control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were 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 splenic leukocytes. 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, washed with Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were washed and then permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with PE Rat Anti-Mouse CD4 antibody Cat. No. 553049 and with either no antibody BD Horizon RB705 Autofluorescence Control; Left Plot or BD Horizon RB705 Rat Anti-Mouse TNF antibody Cat. No. 570733/570734; 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 Autofluorescence control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were 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 splenic leukocytes. 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, washed with Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 . The cells were washed and then permeabilized with and stained in BD Perm/Wash Buffer Cat. No. 554723 with PE Rat Anti-Mouse CD4 antibody Cat. No. 553049 and with either no antibody BD Horizon RB705 Autofluorescence Control; Left Plot or BD Horizon RB705 Rat Anti-Mouse TNF antibody Cat. No. 570733/570734; 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 Autofluorescence control staining versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Cell Analyzer System and FlowJo Software. Data shown on this Technical Data Sheet are not lot specific.