Lane 1: Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using an antibody which recognized an upstream epitope of TBC1D4. Lane 2: Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using an antibody which recognized an upstream epitope of TBC1D4. Lane 3: Detection of Human TBC1D4 by Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using ab134749 at 6 µg/mg lysate for IP. Lane 4: Immunoprecipitation from HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), using Control IgG. Subsequent Western blot detection used ab134749 at 1 µg/ml. Western Blot Gel 4-8% Developed using the ECL technique with an exposure time of 3 seconds.