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Abstract for HaloTag®: A Novel Technology for Cell Analysis
and Protein Immobilization
Chad Zimprich1, Natasha Karassina1, Randy Learish1, Soshana
Behrstock1, Nidhi Nath1, Aldis Darzins1, Marjeta Urh1, Keith V. Wood1,
Georgyi V. Los1, Mark McDougall2 and Dieter Klaubert2
1Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711
2Promega Biosciences, Inc., 277 Granada Drive, San Luis Obispo, CA
93401
The HaloTag® Interchangeable Labeling Technology is a novel tool
for (1) imaging live or fixed mammalian cells that express the
HaloTag® Protein or protein fusions, (2) analyzing
post-translational modification of labeled proteins, and (3)
capturing and immobilizing protein fusions and protein complexes
generated in living cells or in vitro. The technology is based on
the efficient formation of a covalent bond between an engineered
reporting protein and its specific ligand, either in living cells,
in solution, or on a solid support. The reporter protein is a
genetically engineered derivative of a catalytically inactive
hydrolase designed to form a stable, covalent bond to synthetic
ligands. The ligands are small chemical tags capable of carrying a
variety of functionalities, such as fluorescent labels,
environmental sensors, affinity handles, or attachments to a solid
phase. The covalent bond forms rapidly under general physiological
conditions, is highly specific, and essentially irreversible. The
open architecture of the technology ensures interchangeability of
ligands, thereby facilitating a variety of functional studies
(including imaging at different wavelengths and temporal or spatial
separation of protein pools) without requiring changes to the
underlying genetic construct. The HaloTag® Technology complements
existing methods and provides new options for cellular analysis and
protein immobilization. |
