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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 1. Cell-free expression of VMAT2.

Fig 1

A. Native VMAT2 (lane 1), VMAT2 harboring silent mutations (lanes 2 and 3) and synthetic VMAT2 (lane 4) were synthesized by using the rapid-translation system 100 E. coli HY kit. Reaction was supplemented with 35S-Methionine. Samples were separated on SDS-PAGE and radioactive bands were visualized with a FLA-3000 Phosphor-Imager (Fujifilm, Tokyo). B. His tagged VMAT2 (lane 1 N' tag, lane 2 C' tag) was synthesized using the rapid-translation system 100 Wheat Germ HY kit. Reaction was supplemented with 35S-Methionine. Samples were separated on SDS-PAGE and radioactive bands were visualized with a FLA-3000 Phosphor-Imager. C. EAAC1-His synthesized in the rapid-translation system 100 E. coli HY kit was incubated with Ni-NTA beads. After purification and elution from the beads, sample (equivalent to half of the original reaction) was separated by SDS-PAGE and detected by Coomassie blue staining.