ICC/IF image of ab36708 stained Hek293 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 (ab36708, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (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-SCRIBBLE antibody (ab36708) at 1/5000 dilutionLane 1 : Whole cell lysate prepared from prostate cancer cellsLane 2 : Whole cell lysate prepared from prostate cancer cellsLane 3 : Whole cell lysate prepared from prostate cancer cellsLysates/proteins at 40 µg per lane.SecondaryHRP conjugated goat anti-rabbit polyclonal at 1/10000 dilutiondeveloped using the ECL technique