
563376 ·
BDB Bioscience · Cat: 563376
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Description
Two-color flow cytometric analysis of IFN-gamma 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 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553048/553049/561837 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse IFN-gamma antibody Cat. No. 563376; Right Panel by using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the correlated expression of IFN-gamma 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 LSR II Flow Cytometer System. Two-color flow cytometric analysis of IFN-gamma 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 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553048/553049/561837 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse IFN-gamma antibody Cat. No. 563376; Right Panel by using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the correlated expression of IFN-gamma 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 LSR II Flow Cytometer System. Two-color flow cytometric analysis of IFN-gamma 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 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553048/553049/561837 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse IFN-gamma antibody Cat. No. 563376; Right Panel by using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the correlated expression of IFN-gamma 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 LSR II Flow Cytometer System. Show More Two-color flow cytometric analysis of IFN-gamma 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 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553048/553049/561837 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse IFN-gamma antibody Cat. No. 563376; Right Panel by using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the correlated expression of IFN-gamma 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 LSR II Flow Cytometer System. Show More Two-color flow cytometric analysis of IFN-gamma 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 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553048/553049/561837 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse IFN-gamma antibody Cat. No. 563376; Right Panel by using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the correlated expression of IFN-gamma 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 LSR II Flow Cytometer System. Two-color flow cytometric analysis of IFN-gamma 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 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 PE Rat Anti-Mouse CD4 antibody Cat. No. 553048/553049/561837 and either BD Horizon BV421 Rat IgG1, kappa Isotype Control Cat. No. 562868; Left Panel or BD Horizon BV421 Rat Anti-Mouse IFN-gamma antibody Cat. No. 563376; Right Panel by using the BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the correlated expression of IFN-gamma 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 LSR II Flow Cytometer System.