FluoroTectâ„¢ GreenLys in vitro Translation Labeling System Technical Bulletin
Instructions for Use of Product(s)
L5001
Literature # TB285
The FluoroTect™ GreenLys 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™ GreenLys 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.
Summary of Change
The following change was made to the 12/17 revision of this document:
Section 4.B was updated to include details on instrumentation.
Printed in USA. Revised 12/17.