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Cell Surface HaloTag® Technology: Spatial Separation and Bidirectional Trafficking of Proteins
Soshana Svendsen, Ph.D.1, Chad Zimprich,
B.S.1, Mark G. McDougall, Ph.D.2, Dieter H.
Klaubert, Ph.D.2, Georgyi V. Los, Ph.D.1
1Promega Corporation; 2Promega Biosciences
The ability to specifically label proteins within living cells can provide
important information about protein dynamics and function. Here we use a
technology to covalently tether fluorophores with different wavelengths
to the specially designed HaloTag® reporter protein. We have achieved
surface expression of the HaloTag® reporter protein by fusing it to a
truncated integrin. In addition, we have developed a novel membrane-impermeant
fluorophore. By using differently colored cell-impermeant and permeant
fluorophores, we show spatial separation of membrane and intracellular
protein pools, respectively. The truncated integrin protein, labeled
with distinguishable fluorophores, can be followed in real time to study
translocation of surface and intracellular protein pools. Using a
HaloTag®-integrin fusion protein, we have shown the HaloTag® technology
to be a powerful tool to study spatial separation and real-time
translocation of protein pools in
live cells.
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