ab83036 stained PC3 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 ab83036 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
All lanes : Anti-AHA1 antibody (ab83036)Lane 1 : Cell lysates prepared from A431 cellsLane 2 : Cell lysates prepared from A549 cellsLane 3 : Cell lysates prepared from HCT116 cellsLane 4 : Cell lysate prepared from Hela cellsLane 5 : Cell lysate prepared from HEK293 cellsLane 6 : Cell lysates prepared from HepG2 cellsLane 7 : Cell lysates prepared from HL-60 cellsLane 8 : Cell lysates prepared from HUVEC cellsLane 9 : Cell lysates prepared from Jurkat cellsLane 10 : Cell lysates prepared from MCF7 cellsLane 11 : Cell lysates prepared from PC3 cellsLane 12 : Cell lysates prepared from T98G cellsLane 13 : Lysate prepared from rat brainLysates/proteins at 10 µg per lane.