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Going Green

Researchers routinely prepare proteins in test tubes using the in vitro translation (IVT) reaction.1 These reactions are fast, efficient, and reasonably cost-effective. They are also highly flexible. Proteins can be synthesized either from RNA transcribed in vitro (uncoupled), or from plasmid DNA via a coupled transcription/translation system. In addition, the proteins are easily labeled with [35S]-methionine or biotinylated lysine simply by addition of the labeling reagent to the reaction. Unfo

Jeffrey Perkel
Researchers routinely prepare proteins in test tubes using the in vitro translation (IVT) reaction.1 These reactions are fast, efficient, and reasonably cost-effective. They are also highly flexible. Proteins can be synthesized either from RNA transcribed in vitro (uncoupled), or from plasmid DNA via a coupled transcription/translation system. In addition, the proteins are easily labeled with [35S]-methionine or biotinylated lysine simply by addition of the labeling reagent to the reaction. Unfortunately, both of these labeling options have drawbacks. Radiolabeled proteins, separated on SDS-polyacrylamide gels, can only be visualized after the gels have been dried and exposed to film. Likewise, biotinylated proteins must be resolved electrophoretically, blotted to nitrocellulose, and detected via Western blot analysis using streptavidin conjugates.

Now a third labeling option is available, which promises to alleviate the shortcomings of other methods. Madison, Wis.-based Promega Corp.'s FluoroTect™ GreenLys in vitro translation labeling system is a BODIPY...

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