ICC/IF image of ab80181 stained PC12 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 (ab80181, 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.
ab80181, at a 1/50 dilution, staining PAK3 in paraffin embedded human brain tissue by Immunohistochemistry in the absence (left image) or presence (right image) of the immunizing phospho peptide.
All lanes : Anti-PAK3 (phospho S154) antibody (ab80181) at 1/500 dilutionLane 1 : rat heart cell extractsLane 2 : rat heart cell extracts with immunizing phospho peptide at 10 µgLysates/proteins at 30 µg per lane.