ab40754 staining cyclin D1 in MCF7 cells treated with KN-93 (water soluble) (ab120980), by ICC/IF. Decrease in cyclin D1 expression correlates with increased concentration of KN-93 (water soluble), as described in literature.The cells were incubated at 37°C for 24 hours in media containing different concentrations of ab120980 (KN-93 (water soluble)) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab40754 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.