BD PharmingenTM Alexa Fluor 488 Rat Anti-Mouse CD274

Description

Two-color flow cytometric analysis of CD274 expression on resting and activated mouse splenocytes. BALB/c mouse splenic leucocytes were either not activated Upper Plots or activated Lower Plots by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 antibody Cat. No. 553057 for 3 days 37C . The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were then stained with BD Horizon BV421 Rat Anti-Mouse CD4 antibody Cat. No. 562891 and either Alexa Fluor 488 Rat IgG2a, kappa Isotype Control Cat. No. 557676; Left Plots or BD Horizon Alexa Fluor 488 Rat Anti-Mouse CD274 antibody Cat. No. 566864; Right Plots at 0.25 mug/test. BD Via-Probe Cell Viability 7-AAD Solution Cat. No. 555815/555816 was added to cells right before analysis. Two-color flow cytometric contour plots showing the correlated expression of CD274 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light- scatter characteristics of viable 7-AAD-negative leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Two-color flow cytometric analysis of CD274 expression on resting and activated mouse splenocytes. BALB/c mouse splenic leucocytes were either not activated Upper Plots or activated Lower Plots by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 antibody Cat. No. 553057 for 3 days 37C . The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were then stained with BD Horizon BV421 Rat Anti-Mouse CD4 antibody Cat. No. 562891 and either Alexa Fluor 488 Rat IgG2a, kappa Isotype Control Cat. No. 557676; Left Plots or BD Horizon Alexa Fluor 488 Rat Anti-Mouse CD274 antibody Cat. No. 566864; Right Plots at 0.25 mug/test. BD Via-Probe Cell Viability 7-AAD Solution Cat. No. 555815/555816 was added to cells right before analysis. Two-color flow cytometric contour plots showing the correlated expression of CD274 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light- scatter characteristics of viable 7-AAD-negative leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific. Show More Two-color flow cytometric analysis of CD274 expression on resting and activated mouse splenocytes. BALB/c mouse splenic leucocytes were either not activated Upper Plots or activated Lower Plots by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 antibody Cat. No. 553057 for 3 days 37C . The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody Mouse BD Fc Block Cat. No. 553141/553142 . The cells were then stained with BD Horizon BV421 Rat Anti-Mouse CD4 antibody Cat. No. 562891 and either Alexa Fluor 488 Rat IgG2a, kappa Isotype Control Cat. No. 557676; Left Plots or BD Horizon Alexa Fluor 488 Rat Anti-Mouse CD274 antibody Cat. No. 566864; Right Plots at 0.25 mug/test. BD Via-Probe Cell Viability 7-AAD Solution Cat. No. 555815/555816 was added to cells right before analysis. Two-color flow cytometric contour plots showing the correlated expression of CD274 or Ig Isotype control staining versus CD4 were derived from gated events with the forward and side light- scatter characteristics of viable 7-AAD-negative leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa Cell Analyzer System and FlowJo software. Data shown on this Technical Data Sheet are not lot specific.