ICC/IF image of ab22835 stained HeLa 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 (ab22835, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. 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.
Anti-EYA3 antibody (ab22835) at 0.1 µg/ml + HepG2 cell lysate with 5% skim milk in PBSSecondaryHRP conjugated anti-rabbit IgG at 1/50000 dilution
Immunohistochemistry detection of EYA3 in paraffin embedded human skeletal muscle tissue section. ab22835 was used at 4 ug/ml. Arrows indicate positively labelled muscle cells.