All lanes : Anti-WDR77 antibody (ab66299) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell LysateLane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell LysateLane 8 : U2OS (Human osteosarcoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab66299 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 (ab66299, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive result in 100% methanol fixed (5 min) HepG2, MCF-7 cells at 1µg/ml, and in 4% pfa fixed (10 min) HeLa, HepG2, MCF-7 cells at 1µg/ml.