ICC/IF image of ab84666 stained MCF-7 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 ab84666 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Western blot - NMT1 antibody (ab84666)
Anti-NMT1 antibody (ab84666) at 1 µg/ml (in 5% skim milk / PBS buffer) + HepG2 cell lysate at 10 µg
Secondary HRP conjugated anti-Rabbit IgG at 1/50000 dilution
Predicted band size : 57 kDa Observed band size : 57 kDa Additional bands at : 37 kDa,50 kDa. We are unsure as to the identity of these extra bands.