Overlay histogram showing U-2 OS cells using 040735 (green line). The cells were fixed with 2% paraformaldehyde (10min) and then permeabilized with 90% methanol for 10min. The cells were then icubated in 2% BSA to block non-specific protein-protein interactions followed by 040735 (1:25) for 60 min at 37ºC. The secondary antibody used was IgG goat-anti-rabbit, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1:200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was rabbit IgG1 (1ug/1x10e6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.