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. Author manuscript; available in PMC: 2011 Oct 1.
Published in final edited form as: Protein Expr Purif. 2010 Jun 8;73(2):152–160. doi: 10.1016/j.pep.2010.06.001

Fig 4. Expression and purification of VMAT2 expressed in insect cells.

Fig 4

A. Sf9 cells were infected using the baculovirus FBG-VMAT2 vector (lanes 2 and 4). Samples were analyzed by SDS-PAGE and transferred onto PVDF membrane. Western blot was performed using anti-His (lanes 1–2) or anti HA (lanes 3–4) antibodies. Lanes 1 and 3 are un-infected control cells. B. Western analysis using anti-His antibody of Sf9 expressed native VMAT2 (lane 1) and GlyQ in which putative glycosylation sites have been mutated, as described under Material and Methods. C. Purification of Sf9 expressed native VMAT2. Sf9 membranes from 800 ml cells infected using FBG-VMAT2 vector were solubilized (lane 1, 0.5 μl from a total of 60 ml solution) and loaded onto NiNTA column (lane 2 unbound material, same amount as lane 1) and eluted with imidazole (lane 3, 5 μl from a total of 6 ml solution). rVMAT2 was further purified on a ConA column as described (lane 4, 2 μl from a total of 1.5 ml solution). Samples were analyzed on SDS-PAGE and stained with coomassie.