Multicolor in vitro translation View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2003-09

AUTHORS

Giovanni Traverso, Frank Diehl, Robin Hurst, Anthony Shuber, Duncan Whitney, Constance Johnson, Bernard Levin, Kenneth W Kinzler, Bert Vogelstein

ABSTRACT

In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized by detection of labeled amino acids incorporated during translation. The original strategy of incorporating radioactively labeled amino acids, such as [35S]methionine or [14C]leucine, has been superseded by the addition of antigenic tags or the incorporation of biotin-labeled or BODIPY-FL-labeled amino acids. Such nonradioactive tags are easier to visualize after translation and do not pose a radiation hazard. Among the nonradioactive tags, BODIPY-FL-lysine offers the advantage that proteins that have incorporated this amino acid can be directly visualized after gel electrophoresis. We show here that multiple fluorophores introduced into proteins can considerably extend their usefulness, particularly for the comparison of in vitro-translated proteins from related sources. This technology can be applied in various situations, including the simplified detection of rare truncating mutations in clinical samples from cancer patients. More... »

PAGES

1093-1097

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt857

DOI

http://dx.doi.org/10.1038/nbt857

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1023436180

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/12910244


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