BD PhosflowTM PE Mouse anti-IkappaBalpha

Description

Flow cytometric analysis of IkappaBalpha expression. LEFT: IkappaBalpha expression in HeLa cells. HeLa cells ATCC CCL-2 were either treated shaded histogram with 20 ng/mL recombinant human TNF Cat. No. 554618 for 10 min at 37C or untreated open histogram . The cells were fixed BD Cytofix Fixation Buffer, Cat. No. 554655 for 10 minutes at 37C, then permeabilized BD Phosflow Perm Buffer II, Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 . The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. RIGHT: IkappaBalpha expression in human peripheral blood T cells. Human whole blood collected with heparin was either unstimulated open histogram or stimulated shaded histogram with 400 nM PMA plus 250 ng/mL ionomycin Sigma, Cat. No. I-0634 for 15 min at 37C. The erythrocytes were lysed and the leukocytes were fixed with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . The leukocytes were permeabilized with BD Phosflow Perm Buffer II Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 and BD Horizon V450 Mouse Anti-Human CD3 Cat. No. 560365 antibodies. The fluorescence histograms were derived from human CD3-positive T cell-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II flow cytometry system. Flow cytometric analysis of IkappaBalpha expression. LEFT: IkappaBalpha expression in HeLa cells. HeLa cells ATCC CCL-2 were either treated shaded histogram with 20 ng/mL recombinant human TNF Cat. No. 554618 for 10 min at 37C or untreated open histogram . The cells were fixed BD Cytofix Fixation Buffer, Cat. No. 554655 for 10 minutes at 37C, then permeabilized BD Phosflow Perm Buffer II, Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 . The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. RIGHT: IkappaBalpha expression in human peripheral blood T cells. Human whole blood collected with heparin was either unstimulated open histogram or stimulated shaded histogram with 400 nM PMA plus 250 ng/mL ionomycin Sigma, Cat. No. I-0634 for 15 min at 37C. The erythrocytes were lysed and the leukocytes were fixed with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . The leukocytes were permeabilized with BD Phosflow Perm Buffer II Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 and BD Horizon V450 Mouse Anti-Human CD3 Cat. No. 560365 antibodies. The fluorescence histograms were derived from human CD3-positive T cell-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II flow cytometry system. Flow cytometric analysis of IkappaBalpha expression. LEFT: IkappaBalpha expression in HeLa cells. HeLa cells ATCC CCL-2 were either treated shaded histogram with 20 ng/mL recombinant human TNF Cat. No. 554618 for 10 min at 37C or untreated open histogram . The cells were fixed BD Cytofix Fixation Buffer, Cat. No. 554655 for 10 minutes at 37C, then permeabilized BD Phosflow Perm Buffer II, Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 . The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. RIGHT: IkappaBalpha expression in human peripheral blood T cells. Human whole blood collected with heparin was either unstimulated open histogram or stimulated shaded histogram with 400 nM PMA plus 250 ng/mL ionomycin Sigma, Cat. No. I-0634 for 15 min at 37C. The erythrocytes were lysed and the leukocytes were fixed with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . The leukocytes were permeabilized with BD Phosflow Perm Buffer II Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 and BD Horizon V450 Mouse Anti-Human CD3 Cat. No. 560365 antibodies. The fluorescence histograms were derived from human CD3-positive T cell-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II flow cytometry system. Show More Flow cytometric analysis of IkappaBalpha expression. LEFT: IkappaBalpha expression in HeLa cells. HeLa cells ATCC CCL-2 were either treated shaded histogram with 20 ng/mL recombinant human TNF Cat. No. 554618 for 10 min at 37C or untreated open histogram . The cells were fixed BD Cytofix Fixation Buffer, Cat. No. 554655 for 10 minutes at 37C, then permeabilized BD Phosflow Perm Buffer II, Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 . The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. RIGHT: IkappaBalpha expression in human peripheral blood T cells. Human whole blood collected with heparin was either unstimulated open histogram or stimulated shaded histogram with 400 nM PMA plus 250 ng/mL ionomycin Sigma, Cat. No. I-0634 for 15 min at 37C. The erythrocytes were lysed and the leukocytes were fixed with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . The leukocytes were permeabilized with BD Phosflow Perm Buffer II Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 and BD Horizon V450 Mouse Anti-Human CD3 Cat. No. 560365 antibodies. The fluorescence histograms were derived from human CD3-positive T cell-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II flow cytometry system. Show More Flow cytometric analysis of IkappaBalpha expression. LEFT: IkappaBalpha expression in HeLa cells. HeLa cells ATCC CCL-2 were either treated shaded histogram with 20 ng/mL recombinant human TNF Cat. No. 554618 for 10 min at 37C or untreated open histogram . The cells were fixed BD Cytofix Fixation Buffer, Cat. No. 554655 for 10 minutes at 37C, then permeabilized BD Phosflow Perm Buffer II, Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 . The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. RIGHT: IkappaBalpha expression in human peripheral blood T cells. Human whole blood collected with heparin was either unstimulated open histogram or stimulated shaded histogram with 400 nM PMA plus 250 ng/mL ionomycin Sigma, Cat. No. I-0634 for 15 min at 37C. The erythrocytes were lysed and the leukocytes were fixed with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . The leukocytes were permeabilized with BD Phosflow Perm Buffer II Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 and BD Horizon V450 Mouse Anti-Human CD3 Cat. No. 560365 antibodies. The fluorescence histograms were derived from human CD3-positive T cell-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II flow cytometry system. Flow cytometric analysis of IkappaBalpha expression. LEFT: IkappaBalpha expression in HeLa cells. HeLa cells ATCC CCL-2 were either treated shaded histogram with 20 ng/mL recombinant human TNF Cat. No. 554618 for 10 min at 37C or untreated open histogram . The cells were fixed BD Cytofix Fixation Buffer, Cat. No. 554655 for 10 minutes at 37C, then permeabilized BD Phosflow Perm Buffer II, Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 . The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. RIGHT: IkappaBalpha expression in human peripheral blood T cells. Human whole blood collected with heparin was either unstimulated open histogram or stimulated shaded histogram with 400 nM PMA plus 250 ng/mL ionomycin Sigma, Cat. No. I-0634 for 15 min at 37C. The erythrocytes were lysed and the leukocytes were fixed with BD Phosflow Lyse/Fix Buffer Cat. No. 558049 . The leukocytes were permeabilized with BD Phosflow Perm Buffer II Cat. No. 558052 on ice for at least 30 minutes, and then stained with PE Mouse anti-IkappaBalpha Cat. No. 560818 and BD Horizon V450 Mouse Anti-Human CD3 Cat. No. 560365 antibodies. The fluorescence histograms were derived from human CD3-positive T cell-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II flow cytometry system.