ICC/IF image of ab39953 stained mouse embryonic stem cells. The cells were 4% PFA 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 (ab39953, 5µg/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).
All lanes : Anti-Nodal antibody (ab39953) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 3 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 4 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (ab27193)Lane 5 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate Lane 6 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate Lysates/proteins at 20 µg per lane.SecondaryGoat polyclonal to rabbit IgG -H&L - pre adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.