FluoroTect™ Green_Lys in vitro Translation Labeling System Technical...

Literature # TB285

The FluoroTect™ Green_Lys in vitro Translation Labeling System allows fluorescent labeling of in vitro translation products through the use of a modified charged lysine transfer RNA labeled with the fluorophore BODIPY®-FL. Using this system, fluorescently labeled lysine residues are incorporated into nascent proteins during translation, eliminating the requirement for labeling with [³⁵S] methionine or other radioactive amino acids. The fluorescent lysine is added to the translation reaction as a charged epsilon-labeled fluorescent lysine-tRNA complex (FluoroTect™ Green_Lys tRNA) rather than a free amino acid.

Synthesized proteins are resolved by conventional SDS-PAGE analysis. Following gel electrophoresis, detection of labeled proteins is accomplished in 2–5 minutes directly “in-gel” using a laser-based fluorescent scanner. This eliminates any requirements for protein gel manipulation associated with the use of radioactive-labeled amino acids, such as fixing/drying or overnight exposure to X-ray film. The convenience of "in-gel" detection also avoids the time-consuming electroblotting and detection steps of conventional nonisotopic detection systems.