ICC/IF image of ab134756 stained Panc-1 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 ab134756 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
All lanes : Anti-PI4 kinase beta antibody (ab134756) at 0.1 µg/mlLane 1 : Whole cell lysate from HeLa cells at 50 µgLane 2 : Whole cell lysate from HeLa cells at 15 µgLane 3 : Whole cell lysate from 293T cells at 50 µgLane 4 : Whole cell lysate from Jurkat cells at 50 µgdeveloped using the ECL technique
Lane 1: Detection of Human PI4 kinase beta by Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using ab134756 at 6 µg/mg lysate for IP. Lane 2: Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using an antibody which recognized a downstream epitope of PI4 kinase beta. Lane 3: Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using Control IgG. Subsequent Western blot detection used ab134756 at 1 µg/ml. Western Blot Gel 4-8% Developed using the ECL technique with an exposure time of 3 seconds.