ICC/IF image of ab96548 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 (ab96548, 10µg/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.
All lanes : Anti-GTPase HRAS antibody (ab96548) at 1 µg/mlLane 1 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µgLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg/mlSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
developed using the ECL techniquePerformed under reducing conditions.