BD PharmingenTM Purified NA/LE Mouse Anti-Mouse CD28

Description

Flow cytometric analysis of CD28 expression on mouse splenocytes. BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were then stained with either Purified Mouse IgG1, kappa Isotype Control Cat. No. 556648; dashed line histogram or Purified NA/LE Mouse Anti-Mouse CD28 antibody Cat. No. 566883; solid line histogram at 1 mug/test followed by Biotin Rat Anti-Mouse IgG1 antibody Cat. No. 553441 and then PE Streptavidin Cat. No. 554061 . The fluorescence histogram showing CD28 expression or Ig isotype control staining was derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Flow cytometric analysis of proliferating mouse splenocytes stimulated with either Anti-CD3 plus Anti-CD28 antibodies or Anti-CD28 antibody alone. C57BL/6 mouse spleen cells were treated with BD Pharm Lyse Lysing Buffer Cat. No. 555899 to lyse erythrocytes. The splenic leucocytes were washed and labeled with BD Horizon CFSE 5 muM; Cat. No. 565082 for 10 minutes at 37C. The cells were washed twice and then cultured with either Purified NA/LE Hamster Anti-Mouse CD3e 10 mug/ml plate-bound; Cat. No. 553057 and Purified NA/LE Mouse Anti-Mouse CD28 1 mug/ml soluble; Cat. No. 566883 antibodies Left Plot or with Purified NA/LE Mouse Anti-Mouse CD28 antibody 1 mug/ml soluble alone Right Plot . After 3 days, the cells were harvested, washed, and stained with BD Horizon BUV395 Rat Anti-Mouse CD4 antibody Cat. No. 563790 . BD Pharmingen DAPI Solution Cat. No. 564907 was added to samples prior to acquisition to exclude non-viable cells. Two-color flow cytometric dot plots showing the correlated expression of CFSE staining versus CD4 levels were derived from gated events with the forward and side light-scatter characteristics of viable DAPI-negative lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Flow cytometric analysis of CD28 expression on mouse splenocytes. BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were then stained with either Purified Mouse IgG1, kappa Isotype Control Cat. No. 556648; dashed line histogram or Purified NA/LE Mouse Anti-Mouse CD28 antibody Cat. No. 566883; solid line histogram at 1 mug/test followed by Biotin Rat Anti-Mouse IgG1 antibody Cat. No. 553441 and then PE Streptavidin Cat. No. 554061 . The fluorescence histogram showing CD28 expression or Ig isotype control staining was derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Show More Flow cytometric analysis of proliferating mouse splenocytes stimulated with either Anti-CD3 plus Anti-CD28 antibodies or Anti-CD28 antibody alone. C57BL/6 mouse spleen cells were treated with BD Pharm Lyse Lysing Buffer Cat. No. 555899 to lyse erythrocytes. The splenic leucocytes were washed and labeled with BD Horizon CFSE 5 muM; Cat. No. 565082 for 10 minutes at 37C. The cells were washed twice and then cultured with either Purified NA/LE Hamster Anti-Mouse CD3e 10 mug/ml plate-bound; Cat. No. 553057 and Purified NA/LE Mouse Anti-Mouse CD28 1 mug/ml soluble; Cat. No. 566883 antibodies Left Plot or with Purified NA/LE Mouse Anti-Mouse CD28 antibody 1 mug/ml soluble alone Right Plot . After 3 days, the cells were harvested, washed, and stained with BD Horizon BUV395 Rat Anti-Mouse CD4 antibody Cat. No. 563790 . BD Pharmingen DAPI Solution Cat. No. 564907 was added to samples prior to acquisition to exclude non-viable cells. Two-color flow cytometric dot plots showing the correlated expression of CFSE staining versus CD4 levels were derived from gated events with the forward and side light-scatter characteristics of viable DAPI-negative lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Show More Flow cytometric analysis of CD28 expression on mouse splenocytes. BALB/c mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were then stained with either Purified Mouse IgG1, kappa Isotype Control Cat. No. 556648; dashed line histogram or Purified NA/LE Mouse Anti-Mouse CD28 antibody Cat. No. 566883; solid line histogram at 1 mug/test followed by Biotin Rat Anti-Mouse IgG1 antibody Cat. No. 553441 and then PE Streptavidin Cat. No. 554061 . The fluorescence histogram showing CD28 expression or Ig isotype control staining was derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Flow cytometric analysis of proliferating mouse splenocytes stimulated with either Anti-CD3 plus Anti-CD28 antibodies or Anti-CD28 antibody alone. C57BL/6 mouse spleen cells were treated with BD Pharm Lyse Lysing Buffer Cat. No. 555899 to lyse erythrocytes. The splenic leucocytes were washed and labeled with BD Horizon CFSE 5 muM; Cat. No. 565082 for 10 minutes at 37C. The cells were washed twice and then cultured with either Purified NA/LE Hamster Anti-Mouse CD3e 10 mug/ml plate-bound; Cat. No. 553057 and Purified NA/LE Mouse Anti-Mouse CD28 1 mug/ml soluble; Cat. No. 566883 antibodies Left Plot or with Purified NA/LE Mouse Anti-Mouse CD28 antibody 1 mug/ml soluble alone Right Plot . After 3 days, the cells were harvested, washed, and stained with BD Horizon BUV395 Rat Anti-Mouse CD4 antibody Cat. No. 563790 . BD Pharmingen DAPI Solution Cat. No. 564907 was added to samples prior to acquisition to exclude non-viable cells. Two-color flow cytometric dot plots showing the correlated expression of CFSE staining versus CD4 levels were derived from gated events with the forward and side light-scatter characteristics of viable DAPI-negative lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa X-20 Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific.