All lanes : Anti-eIF4GII antibody (ab40996) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell LysateLane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 4 : U2OS (Human osteosarcoma cell line) Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.
ICC/IF image of ab40996 stained human MCF7 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab40996, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and HepG2 cells.