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Abstract for the FluoroTect™ GreenLys in vitro Translation Labeling
System
The FluoroTect™ GreenLys in vitro Translation Labeling System
allows the 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 the 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
non-isotopic detection systems. |
