ICC/IF image of ab103063 stained HeLa 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 ab103063 at 1µg/ml 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-HIF-1-alpha antibody (ab103063) at 2 µg/mlLane 1 : Hela-Vehicle treated (Negative Control) Whole Cell Lysate (ab116321)Lane 2 : Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322)Lysates/proteins at 40 µg per lane.SecondaryAnti-Rabbit (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.