ICC/IF image of ab82749 stained U2OS cells. The cells were 100% methanol fixed (5 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 ab82749 at Undiluted (Used Neat) overnight at +4°C. The secondary antibody (pseudo-colored green) was Dylight® 488 goat anti- mouse (ab98674) IgM preadsorbed, used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.