Overlay histogram showing Jurkat cells stained with antibody (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then incubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (antibody, 1:25 dilution) for 60 min at 37°C. The secondary antibody used was Goat-Anti-mouse IgG, DyLight 488 Conjugated Highly Cross-Adsorbed) at 1:400 dilution for 40 min at 37 ° C. Isotype control antibody (blue line) was mouse IgG (1ug/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.