| Labeling Of Proteins In Living Cells - The SNAP-tag Technology |
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Presentation: 5B01 Session: Imaging and High Content Screening: Valuable Tools Equally Embraced by Academia & Drug Screeners - Part 2Andreas Brecht, Jan Barnikow, Maik Kindermann, Markus Schwab and Ron Tynes, Covalys Biosciences AG Presenting Author: Andreas Brecht, Covalys Biosciences AG - Switzerland Assays on cellular protein dynamics and function rely crucially on labeling of proteins in living cells. SNAP-tag is a novel protein-tag which labels itself covalently with a wide range of fluorescent or affinity substrates. Highly specific labeling results in strictly one label per SNAP-tag. Typically a 15 min incubation of living cells with the cell-permeable label is followed by a brief washout of nonreacted label. SNAP-tag fusion proteins have been successfully expressed and labeled in different cell compartments and in various mammalian cell-lines. Independence of expression and labeling allows for 'pulse-chase' applications. Exclusive labeling of membrane receptors on the extracellular side of the membrane, without getting a signal from immature intracellular receptors, is achieved using non cell-permeable labels. SNAP-tag labels tolerate typical fixation conditions, offer a wide range of colors, are easy to use, and come without licensing requirements, making them an attractive alternative to fluorescent proteins for screening applications. |