Skip to main content
. Author manuscript; available in PMC: 2016 May 9.
Published in final edited form as: Biochem J. 2010 Jul 15;429(2):391–401. doi: 10.1042/BJ20100336

Figure 4. Sec22b pulls down a ~110 kDa protein complex from post-docking VTV–Golgi complexes.

Figure 4

VTVs (150 μg) were incubated at 4 °C with hepatic cis-Golgi (300 μg), cytosol (500 μg of protein) and without Mg2+-ATP. The VTV–Golgi complexes were isolated on a sucrose step gradient. The VTV–Golgi complexes were solubilized in 2 % (v/v) Triton X-100 and incubated with either pre-immune IgG or anti-Sec22b antibodies bound to agarose beads at 4 °C overnight. The beads were washed and either boiled or not, as indicated, in Laemmli buffer. The proteins were separated by SDS/PAGE (5–15 % gel) and the gel was stained. For further details, see the Experimental section. IP, immunoprecipitation.