Anti cd3 fitc cd8pe cd45percp cd4apc antibodies

Two BD Trucount tubes (A and B) containing a known number of fluorescent beads were consecutively numbered and 50μL of fully mixed anticoagulant whole blood was added. Added 20μL anti-CD3-FITC/CD8PE/CD45PercP/CD4APC antibodies into tube A, 20μL anti-CD3-FITC/CD16+56-PE/CD45-PercP/CD19-APC antibodies were added to B tube (all from BD Biosciences). Mixed the tube contents thoroughly and incubated at 25°C, without light for 15-20 minutes. Then, 450μL 1X FACS hemolysin was added and fully mixed, and incubated at 25°C, dark for 15 minutes. Washed with phosphate buffered saline (PBS) and tested on the machine within 24 hours. For each sample, 15000 cells were obtained for analysis using BD MultiSET software (BD Biosciences).

Two BD Trucount tubes containing a known number of fluorescent beads were serially numbered (A and B), and 50 μl blood samples were added to the tubes by reverse loading. Then, 20 μl of anti-CD3-FITC/CD-8PE/CD45PercP/CD4APC antibodies was added to tube A, and 20 μl of CD3FITC/CD16+56-PE/CD45-PercP/CD19-APC antibodies was added to tube B (all from BD Biosciences). The contents of the tubes were mixed and incubated at room temperature for 15–20 min in the dark. Then, 450 μl XFACS hemolysin was added and mixed thoroughly, followed by incubation at room temperature for 15 min in the dark. In total, 15000 cells were obtained for detection in 24 h and analyzed by flow cytometry and MultiSET software (BD Biosciences) (Figure 1(a)).