BD PharmingenTM PE Mouse anti-T-Bet

Description

Flow cytometric analysis of T-bet expression by human peripheral blood lymphocytes. Left and Middle Left Panels: Whole blood was treated with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 to lyse erythrocytes and fix leukocytes. Cells were permeabilized with BD Phosflow Perm/Wash Buffer I Cat. No. 557885; Left Panel or Perm Buffer III Cat. No. 558050; Middle Left Panel . Cells were stained with Alexa Fluor 647 Mouse anti-Human CD3 Cat. No. 557706 and PE Mouse anti-T-Bet Cat. No. 561265 antibodies. Two-color dot plots showing CD3 versus T-bet expression for lymphocytes were generated using a BD FACSCalibur Flow Cytometer. Middle Right Panel: Whole blood was treated with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . Cells were permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 , washed, and stained with PE Mouse anti-T-Bet Cat. No. 561265 , FITC Mouse anti-Human CD45RO Cat. No. 555492 , APC Mouse anti-Human CD45RA Cat. No. 550855 , BD Horizon V450 Mouse anti-Human CD3 Cat. No. 560365 , and PerCP Mouse anti-Human CD4 Cat. No. 347324 antibodies. Two-color contour plots showing CD45RA Top Plot or CD45RO Bottom Plot versus T-bet expression patterns for CD3+CD4+ lymphocytes were generated using a BD FACSCanto II Flow Cytometer. Right Panel: Human peripheral blood mononuclear cells were polarized in culture to generate Th1- shaded histogram or Th2-like open histogram cells. The cells were restimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634; 1 ug/ml . The cells were then fixed, permeabilized, and stained with PE Mouse anti-T-Bet Cat. No. 561265 , V450 Mouse anti-Human CD3 Cat. No. 560366 and PerCP-Cy5.5 Mouse Anti-Human CD4 Cat. No. 560650 using BD Biosciences Protocol for Intracellular Cytokine Staining. The histograms for CD3+CD4+ lymphocytes expressing T-bet were generated using a BD FACSCanto II Flow Cytometer. Flow cytometric analysis of T-bet expression by human peripheral blood lymphocytes. Left and Middle Left Panels: Whole blood was treated with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 to lyse erythrocytes and fix leukocytes. Cells were permeabilized with BD Phosflow Perm/Wash Buffer I Cat. No. 557885; Left Panel or Perm Buffer III Cat. No. 558050; Middle Left Panel . Cells were stained with Alexa Fluor 647 Mouse anti-Human CD3 Cat. No. 557706 and PE Mouse anti-T-Bet Cat. No. 561265 antibodies. Two-color dot plots showing CD3 versus T-bet expression for lymphocytes were generated using a BD FACSCalibur Flow Cytometer. Middle Right Panel: Whole blood was treated with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . Cells were permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 , washed, and stained with PE Mouse anti-T-Bet Cat. No. 561265 , FITC Mouse anti-Human CD45RO Cat. No. 555492 , APC Mouse anti-Human CD45RA Cat. No. 550855 , BD Horizon V450 Mouse anti-Human CD3 Cat. No. 560365 , and PerCP Mouse anti-Human CD4 Cat. No. 347324 antibodies. Two-color contour plots showing CD45RA Top Plot or CD45RO Bottom Plot versus T-bet expression patterns for CD3+CD4+ lymphocytes were generated using a BD FACSCanto II Flow Cytometer. Right Panel: Human peripheral blood mononuclear cells were polarized in culture to generate Th1- shaded histogram or Th2-like open histogram cells. The cells were restimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634; 1 ug/ml . The cells were then fixed, permeabilized, and stained with PE Mouse anti-T-Bet Cat. No. 561265 , V450 Mouse anti-Human CD3 Cat. No. 560366 and PerCP-Cy5.5 Mouse Anti-Human CD4 Cat. No. 560650 using BD Biosciences Protocol for Intracellular Cytokine Staining. The histograms for CD3+CD4+ lymphocytes expressing T-bet were generated using a BD FACSCanto II Flow Cytometer. Show More Flow cytometric analysis of T-bet expression by human peripheral blood lymphocytes. Left and Middle Left Panels: Whole blood was treated with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 to lyse erythrocytes and fix leukocytes. Cells were permeabilized with BD Phosflow Perm/Wash Buffer I Cat. No. 557885; Left Panel or Perm Buffer III Cat. No. 558050; Middle Left Panel . Cells were stained with Alexa Fluor 647 Mouse anti-Human CD3 Cat. No. 557706 and PE Mouse anti-T-Bet Cat. No. 561265 antibodies. Two-color dot plots showing CD3 versus T-bet expression for lymphocytes were generated using a BD FACSCalibur Flow Cytometer. Middle Right Panel: Whole blood was treated with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . Cells were permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 , washed, and stained with PE Mouse anti-T-Bet Cat. No. 561265 , FITC Mouse anti-Human CD45RO Cat. No. 555492 , APC Mouse anti-Human CD45RA Cat. No. 550855 , BD Horizon V450 Mouse anti-Human CD3 Cat. No. 560365 , and PerCP Mouse anti-Human CD4 Cat. No. 347324 antibodies. Two-color contour plots showing CD45RA Top Plot or CD45RO Bottom Plot versus T-bet expression patterns for CD3+CD4+ lymphocytes were generated using a BD FACSCanto II Flow Cytometer. Right Panel: Human peripheral blood mononuclear cells were polarized in culture to generate Th1- shaded histogram or Th2-like open histogram cells. The cells were restimulated with PMA Sigma Cat. No. P-8139 and Ionomycin Sigma Cat. No. I-0634; 1 ug/ml . The cells were then fixed, permeabilized, and stained with PE Mouse anti-T-Bet Cat. No. 561265 , V450 Mouse anti-Human CD3 Cat. No. 560366 and PerCP-Cy5.5 Mouse Anti-Human CD4 Cat. No. 560650 using BD Biosciences Protocol for Intracellular Cytokine Staining. The histograms for CD3+CD4+ lymphocytes expressing T-bet were generated using a BD FACSCanto II Flow Cytometer.