ICC/IF image of ab74504 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 ab74504 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 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.
All lanes : Anti-ZDHHC9 antibody (ab74504) at 1/500 dilutionLane 1 : extracts from COLO cellsLane 2 : extracts from COLO cells with immunising peptide at 10 µgLysates/proteins at 30 µg per lane.
Immunohistochemistry analysis of paraffin-embedded human brain tissue using 1/50 ab74504. Left: untreated sample. Right: sample treated with immunising peptide.