Flow cytometry analysis of CD3 was performed in Jurkat cells. Cells were fixed with 1% paraformaldehyde for 20 mins and blocked in PBS containing 2.5% BSA for 45 minutes at room temperature. Cells were labeled with CD3 Mouse Monoclonal Antibody (MHCD03004, red histogram) at a dilution of 1:20 or a matched isotype control (pink histogram) in PBS. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature. A representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.