Two-color flow cytometric analysis of CXCL16 expression in mouse splenocytes. C57BL/6 mouse splenic leucocytes were cultured in complete tissue culture medium with BD GolgiPlug™ Protein Transport Inhibitor (containing Brefeldin A) [Cat. No. 555029; 5h, 37°C). The cell were harvested, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained with FITC Hamster Anti-Mouse CD3e (Cat. No. 553062/553061/561827), BD Horizon™ BUV395 Rat Anti-Mouse CD19 (Cat. No. 563557/565965), APC Mouse Anti-Mouse NK1.1 antibody (Cat. No. 550627/561117), BD Horizon BV421 Rat Anti-Mouse CD11b (Cat. No. 562605) antibodies and either PE Rat IgG1, κ Isotype Control (Cat. No. 554685; Left Plot) or PE Rat Anti-Mouse CXCL16 antibody (Cat. No. 566740; Right Plot) at 0.5 µg /test. The two-color dot plot showing the correlated expression of CXCL16 (or Ig Isotype control staining) versus CD11b was derived from CD3- CD19- NK1.1- gated events with the forward and side light-scatter characteristics of intact splenocytes. 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.