Overlay histogram showing MCF-7 cells stained with ab72813 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab72813, 0.5µg/1x10⁶ cells) for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 (FITC) (2µg/1x10⁶ cells) for 30 min at 22°C. Acquisition of >5,000 events was performed.

ICC/IF image of ab72813 stained HCT116 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab72813, 1µg/ml) overnight at +4°C. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.