BD PhosflowTM Alexa Fluor 647 Mouse anti-SLP-76 (pY128)

Description

Analysis of SLP-76 pY128 in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were either unstimulated open histograms or stimulated by cross-linking with CD3 Purified NA/LE Mouse anti-Human CD3 mAb, clone UCHT1,Cat. No. 555329 and CD28 Purified NA/LE Mouse anti-Human CD28, clone CD28.2, Cat. No. 555725 on ice for 15 minutes followed by incubation with the 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 histograms . The cells were fixed BD Cytofix Fixation buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-SLP-76 pY128 Cat. No. 558438 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . The left panel displays the upregulated phosphorylation of SLP-76 in activated CD3-positive T lymphocytes, while the right panel displays the lack of upregulated SLP-76 phosphorylation in CD3-negative non-T cells. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. Analysis of SLP-76 pY128 in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were either unstimulated open histograms or stimulated by cross-linking with CD3 Purified NA/LE Mouse anti-Human CD3 mAb, clone UCHT1,Cat. No. 555329 and CD28 Purified NA/LE Mouse anti-Human CD28, clone CD28.2, Cat. No. 555725 on ice for 15 minutes followed by incubation with the 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 histograms . The cells were fixed BD Cytofix Fixation buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-SLP-76 pY128 Cat. No. 558438 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . The left panel displays the upregulated phosphorylation of SLP-76 in activated CD3-positive T lymphocytes, while the right panel displays the lack of upregulated SLP-76 phosphorylation in CD3-negative non-T cells. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. Analysis of SLP-76 pY128 in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were either unstimulated open histograms or stimulated by cross-linking with CD3 Purified NA/LE Mouse anti-Human CD3 mAb, clone UCHT1,Cat. No. 555329 and CD28 Purified NA/LE Mouse anti-Human CD28, clone CD28.2, Cat. No. 555725 on ice for 15 minutes followed by incubation with the 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 histograms . The cells were fixed BD Cytofix Fixation buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-SLP-76 pY128 Cat. No. 558438 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . The left panel displays the upregulated phosphorylation of SLP-76 in activated CD3-positive T lymphocytes, while the right panel displays the lack of upregulated SLP-76 phosphorylation in CD3-negative non-T cells. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. Show More Analysis of SLP-76 pY128 in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were either unstimulated open histograms or stimulated by cross-linking with CD3 Purified NA/LE Mouse anti-Human CD3 mAb, clone UCHT1,Cat. No. 555329 and CD28 Purified NA/LE Mouse anti-Human CD28, clone CD28.2, Cat. No. 555725 on ice for 15 minutes followed by incubation with the 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 histograms . The cells were fixed BD Cytofix Fixation buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-SLP-76 pY128 Cat. No. 558438 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . The left panel displays the upregulated phosphorylation of SLP-76 in activated CD3-positive T lymphocytes, while the right panel displays the lack of upregulated SLP-76 phosphorylation in CD3-negative non-T cells. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. Show More Analysis of SLP-76 pY128 in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were either unstimulated open histograms or stimulated by cross-linking with CD3 Purified NA/LE Mouse anti-Human CD3 mAb, clone UCHT1,Cat. No. 555329 and CD28 Purified NA/LE Mouse anti-Human CD28, clone CD28.2, Cat. No. 555725 on ice for 15 minutes followed by incubation with the 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 histograms . The cells were fixed BD Cytofix Fixation buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-SLP-76 pY128 Cat. No. 558438 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . The left panel displays the upregulated phosphorylation of SLP-76 in activated CD3-positive T lymphocytes, while the right panel displays the lack of upregulated SLP-76 phosphorylation in CD3-negative non-T cells. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. Analysis of SLP-76 pY128 in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were either unstimulated open histograms or stimulated by cross-linking with CD3 Purified NA/LE Mouse anti-Human CD3 mAb, clone UCHT1,Cat. No. 555329 and CD28 Purified NA/LE Mouse anti-Human CD28, clone CD28.2, Cat. No. 555725 on ice for 15 minutes followed by incubation with the 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 histograms . The cells were fixed BD Cytofix Fixation buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-SLP-76 pY128 Cat. No. 558438 and PerCP-Cy5.5 Mouse anti-human CD3 mAb SK7 Cat. No. 340949 . The left panel displays the upregulated phosphorylation of SLP-76 in activated CD3-positive T lymphocytes, while the right panel displays the lack of upregulated SLP-76 phosphorylation in CD3-negative non-T cells. Flow cytometry was performed on a BD FACSCalibur flow cytometry system.