BD PhosflowTM PE Mouse anti-Zap70 (pY292)

Description

LEFT: Analysis of ZAP70 pY292 in activated human T lymphocytes. Human whole blood was either stimulated by cross-linking of CD3 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10-15 minutes at 37oC, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. For data analysis, CD3-positive and -negative lymphocytes were selected by their scatter and staining profiles. The figure displays the CD3-positive T lymphocytes. Up-regulated phosphorylation of ZAP70 pY292 was not observed in the CD3-negative lymphocytes data not shown . RIGHT: Analysis of ZAP70 pY292 in activated human T leukemia cells. Jurkat cells ATCC TIB152 were either stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb CD28.2 Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. LEFT: Analysis of ZAP70 pY292 in activated human T lymphocytes. Human whole blood was either stimulated by cross-linking of CD3 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10-15 minutes at 37oC, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. For data analysis, CD3-positive and -negative lymphocytes were selected by their scatter and staining profiles. The figure displays the CD3-positive T lymphocytes. Up-regulated phosphorylation of ZAP70 pY292 was not observed in the CD3-negative lymphocytes data not shown . RIGHT: Analysis of ZAP70 pY292 in activated human T leukemia cells. Jurkat cells ATCC TIB152 were either stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb CD28.2 Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. LEFT: Analysis of ZAP70 pY292 in activated human T lymphocytes. Human whole blood was either stimulated by cross-linking of CD3 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10-15 minutes at 37oC, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. For data analysis, CD3-positive and -negative lymphocytes were selected by their scatter and staining profiles. The figure displays the CD3-positive T lymphocytes. Up-regulated phosphorylation of ZAP70 pY292 was not observed in the CD3-negative lymphocytes data not shown . RIGHT: Analysis of ZAP70 pY292 in activated human T leukemia cells. Jurkat cells ATCC TIB152 were either stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb CD28.2 Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. Show More LEFT: Analysis of ZAP70 pY292 in activated human T lymphocytes. Human whole blood was either stimulated by cross-linking of CD3 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10-15 minutes at 37oC, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. For data analysis, CD3-positive and -negative lymphocytes were selected by their scatter and staining profiles. The figure displays the CD3-positive T lymphocytes. Up-regulated phosphorylation of ZAP70 pY292 was not observed in the CD3-negative lymphocytes data not shown . RIGHT: Analysis of ZAP70 pY292 in activated human T leukemia cells. Jurkat cells ATCC TIB152 were either stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb CD28.2 Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. Show More LEFT: Analysis of ZAP70 pY292 in activated human T lymphocytes. Human whole blood was either stimulated by cross-linking of CD3 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10-15 minutes at 37oC, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. For data analysis, CD3-positive and -negative lymphocytes were selected by their scatter and staining profiles. The figure displays the CD3-positive T lymphocytes. Up-regulated phosphorylation of ZAP70 pY292 was not observed in the CD3-negative lymphocytes data not shown . RIGHT: Analysis of ZAP70 pY292 in activated human T leukemia cells. Jurkat cells ATCC TIB152 were either stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb CD28.2 Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. LEFT: Analysis of ZAP70 pY292 in activated human T lymphocytes. Human whole blood was either stimulated by cross-linking of CD3 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10-15 minutes at 37oC, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system. For data analysis, CD3-positive and -negative lymphocytes were selected by their scatter and staining profiles. The figure displays the CD3-positive T lymphocytes. Up-regulated phosphorylation of ZAP70 pY292 was not observed in the CD3-negative lymphocytes data not shown . RIGHT: Analysis of ZAP70 pY292 in activated human T leukemia cells. Jurkat cells ATCC TIB152 were either stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb UCHT1 Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb CD28.2 Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 5 minutes shaded histogram , or unstimulated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with PE Mouse anti-ZAP70 pY292 . Flow cytometry was performed on a BD FACSCalibur flow cytometry system.