Description
This system enables fluorescent labeling and detection of in vitro synthesized proteins, based on a lysine-charged tRNA labeled with the fluorophore BODIPY®-FL at the ε position. Fluorescent lysine residues are incorporated into synthesized proteins during in vitro translation reactions, eliminating the need for radioactivity.
Detection of the labeled proteins is accomplished in 2–5 minutes directly "in-gel" by use of a laser-based fluorescent gel scanner. This eliminates any need for protein gel manipulation such as fixing/drying or any safety, regulatory and waste disposal issues associated with the use of radioactively labeled amino acids. The convenience of "in-gel" detection also avoids the time-consuming electroblotting and detection steps of conventional non-isotopic systems.
The modified charged tRNA can be used with a variety of Promega translation systems including: Rabbit Reticulocyte Lysate, T n T® Coupled Transcription/Translation System, Wheat Germ Extract and E. coli S30 Extract.
For more information, see the Protocols & Applications Guide .
Schematic illustrating the incorporation of FluoroTect™ Green Lys -labeled lysine into nascent protein.
