All lanes : Anti-NPL4 antibody (ab101226) at 1 µg/mlLane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate Lane 4 : PC3 (Human prostate cancer cell line) Whole Cell Lysate (ab3954)Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab101226 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 ab101226 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in formaldehyde fixed (4%, 10min) Hek293, HepG2, amd MCF-7 cells at 5ug/ml.