ICC/IF image of ab134218 stained mES 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 (ab134218, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96947, DyLight® 488 goat anti-chicken IgY (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-Oct4 antibody (ab134218) at 1 µg/mlLane 1 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell LysateLane 2 : F9 (Mouse embryonic carcinoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.