ICC/IF image of ab93798 stained HeLa 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 ab93798 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.
All lanes : Anti-MLK4 antibody (ab93798) at 0.04 µg/mlLane 1 : HeLa whole cell lysate at 50 µgLane 2 : HeLa whole cell lysate at 15 µgLane 3 : HeLa whole cell lysate at 5 µgLane 4 : 293T whole cell lysate at 50 µgdeveloped using the ECL technique
Detection of MLK4 in Immunoprecipitates of HeLa whole cell lysate (1 mg for IP, 20% of IP loaded) using ab93798 at 3 µg/mg lysate for IP (Lane 1) and at 1 µg/ml for subsequent Western blot detection. Lane 2 represents control IgG IP. Detection: Chemiluminescence with an exposure time of 3 seconds.