BD PharmingenTM Purified Mouse Anti-Human Chromogranin A

Description

Flow cytometric analysis of Chromogranin A expression in human neuroblastoma cells. SH-SY5Y neuroblastoma cells ATCC CRL-2266 were fixed and permeabilized using BD Cytofix/Cytoperm Fixation/Permeablization Kit Cat. No. 554714 , washed twice with BD Perm/Wash Buffer Cat. No. 554723 , then stained with either Purified Mouse Anti-Human Chromogranin A or Purified Mouse IgG1, kappa Isotype Control Cat. No.555746 monoclonal antibodies followed by PE Goat Anti-Mouse Ig Multiple Adsorption Cat. No. 550589 second-step reagent. The fixed cells were resuspended in stain buffer and acquired on a BD LSR II Flow Cytometry System. The dotted line shows isotype control background staining, while the solid line represents Chromogranin A staining. The overlay histograms were derived from gated events with forward and side light-scatter characteristics of viable cells. Additionally, Purified Mouse Anti-Human Chromogranin A was negative on human embryonic stem ES cells data not shown . Western Blot analysis of Chromogranin A in human SH-SY5Y neuroblastoma lysate. SH-SY5Y neuroblastoma ATCC CRL-2266 cell lysate was probed with Purified Mouse Anti-Human Chromogranin A monoclonal antibody at titrations of 4.0 to 0.007 ug/lane. Data shown in lanes 1,2,3 are two fold dilutions; 0.25ug, 0.125ug, 0.063ug. Chromogranin A is identified as a band of 70 kDa. Immunofluorescent staining of Chromogranin A in human neuroblastoma cells. SH-SY5Y neuroblastoma cells ATCC CRL-2266 were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized with BD Perm/Wash Buffer Cat. No. 554723 and stained with Purified Mouse Anti-Human Chromogranin A monoclonal antibody pseudo-colored green at 1.2 mug/mL. The second-step reagent was Alexa Fluor 488 goat anti-mouse Ig Life Technologies , and counter staining was with DAPI pseudo-colored blue . No Chromogranin A staining was observed on human embryonic stem cells hESC that were imaged similarly data not shown . Also, no staining was observed when SH-SY5Y and hESC were stained with second-step reagent alone data not shown . The image was captured on a BD Pathway 435 Cell Analyzer and merged using BD Attovision Software. BD Phosflow Perm Buffer III Cat. No. 558050 or Triton X-100 are also suitable for permeabilization. Flow cytometric analysis of Chromogranin A expression in human neuroblastoma cells. SH-SY5Y neuroblastoma cells ATCC CRL-2266 were fixed and permeabilized using BD Cytofix/Cytoperm Fixation/Permeablization Kit Cat. No. 554714 , washed twice with BD Perm/Wash Buffer Cat. No. 554723 , then stained with either Purified Mouse Anti-Human Chromogranin A or Purified Mouse IgG1, kappa Isotype Control Cat. No.555746 monoclonal antibodies followed by PE Goat Anti-Mouse Ig Multiple Adsorption Cat. No. 550589 second-step reagent. The fixed cells were resuspended in stain buffer and acquired on a BD LSR II Flow Cytometry System. The dotted line shows isotype control background staining, while the solid line represents Chromogranin A staining. The overlay histograms were derived from gated events with forward and side light-scatter characteristics of viable cells. Additionally, Purified Mouse Anti-Human Chromogranin A was negative on human embryonic stem ES cells data not shown . Show More Western Blot analysis of Chromogranin A in human SH-SY5Y neuroblastoma lysate. SH-SY5Y neuroblastoma ATCC CRL-2266 cell lysate was probed with Purified Mouse Anti-Human Chromogranin A monoclonal antibody at titrations of 4.0 to 0.007 ug/lane. Data shown in lanes 1,2,3 are two fold dilutions; 0.25ug, 0.125ug, 0.063ug. Chromogranin A is identified as a band of 70 kDa. Show More Immunofluorescent staining of Chromogranin A in human neuroblastoma cells. SH-SY5Y neuroblastoma cells ATCC CRL-2266 were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized with BD Perm/Wash Buffer Cat. No. 554723 and stained with Purified Mouse Anti-Human Chromogranin A monoclonal antibody pseudo-colored green at 1.2 mug/mL. The second-step reagent was Alexa Fluor 488 goat anti-mouse Ig Life Technologies , and counter staining was with DAPI pseudo-colored blue . No Chromogranin A staining was observed on human embryonic stem cells hESC that were imaged similarly data not shown . Also, no staining was observed when SH-SY5Y and hESC were stained with second-step reagent alone data not shown . The image was captured on a BD Pathway 435 Cell Analyzer and merged using BD Attovision Software. BD Phosflow Perm Buffer III Cat. No. 558050 or Triton X-100 are also suitable for permeabilization. Show More Flow cytometric analysis of Chromogranin A expression in human neuroblastoma cells. SH-SY5Y neuroblastoma cells ATCC CRL-2266 were fixed and permeabilized using BD Cytofix/Cytoperm Fixation/Permeablization Kit Cat. No. 554714 , washed twice with BD Perm/Wash Buffer Cat. No. 554723 , then stained with either Purified Mouse Anti-Human Chromogranin A or Purified Mouse IgG1, kappa Isotype Control Cat. No.555746 monoclonal antibodies followed by PE Goat Anti-Mouse Ig Multiple Adsorption Cat. No. 550589 second-step reagent. The fixed cells were resuspended in stain buffer and acquired on a BD LSR II Flow Cytometry System. The dotted line shows isotype control background staining, while the solid line represents Chromogranin A staining. The overlay histograms were derived from gated events with forward and side light-scatter characteristics of viable cells. Additionally, Purified Mouse Anti-Human Chromogranin A was negative on human embryonic stem ES cells data not shown . Western Blot analysis of Chromogranin A in human SH-SY5Y neuroblastoma lysate. SH-SY5Y neuroblastoma ATCC CRL-2266 cell lysate was probed with Purified Mouse Anti-Human Chromogranin A monoclonal antibody at titrations of 4.0 to 0.007 ug/lane. Data shown in lanes 1,2,3 are two fold dilutions; 0.25ug, 0.125ug, 0.063ug. Chromogranin A is identified as a band of 70 kDa. Immunofluorescent staining of Chromogranin A in human neuroblastoma cells. SH-SY5Y neuroblastoma cells ATCC CRL-2266 were fixed with BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized with BD Perm/Wash Buffer Cat. No. 554723 and stained with Purified Mouse Anti-Human Chromogranin A monoclonal antibody pseudo-colored green at 1.2 mug/mL. The second-step reagent was Alexa Fluor 488 goat anti-mouse Ig Life Technologies , and counter staining was with DAPI pseudo-colored blue . No Chromogranin A staining was observed on human embryonic stem cells hESC that were imaged similarly data not shown . Also, no staining was observed when SH-SY5Y and hESC were stained with second-step reagent alone data not shown . The image was captured on a BD Pathway 435 Cell Analyzer and merged using BD Attovision Software. BD Phosflow Perm Buffer III Cat. No. 558050 or Triton X-100 are also suitable for permeabilization.