HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein Analysis

@article{citeulike:2870105, abstract = {We have designed a modular protein tagging system that allows different functionalities to be linked onto a single genetic fusion, either in solution, in living cells, or in chemically fixed cells. The protein tag (HaloTag) is a modified haloalkane dehalogenase designed to covalently bind to synthetic ligands (HaloTag ligands). The synthetic ligands comprise a chloroalkane linker attached to a variety of useful molecules, such as fluorescent dyes, affinity handles, or solid surfaces. Covalent bond formation between the protein tag and the chloroalkane linker is highly specific, occurs rapidly under physiological conditions, and is essentially irreversible. We demonstrate the utility of this system for cellular imaging and protein immobilization by analyzing multiple molecular processes associated with NF-ºB-mediated cellular physiology, including imaging of subcellular protein translocation and capture of proteinprotein and proteinDNA complexes.}, address = {Promega Corporation, 2800 Woods Hollow Rd., Madison, Wisconsin 53711, !Promega Biosciences Incorporated, 277 Granada Dr., San Luis Obispo, California 93401, {\S}Present address: National Renewable Energy Laboratory, 1617 Cole Blvd., Golden, Colorado 80401}, author = {Los, Georgyi V. and Encell, Lance P. and Mcdougall, Mark G. and Hartzell, Danette D. and Karassina, Natasha and Zimprich, Chad and Wood, Monika G. and Learish, Randy and Ohana, Rachel F. and Urh, Marjeta and Simpson, Dan and Mendez, Jacqui and Zimmerman, Kris and Otto, Paul and Vidugiris, Gediminas and Zhu, Ji and Darzins, Aldis and Klaubert, Dieter H. and Bulleit, Robert F. and Wood, Keith V. }, citeulike-article-id = {2870105}, doi = {http://dx.doi.org/10.1021/cb800025k}, journal = {ACS Chem. Biol.}, keywords = {fluorescence, fluorescent-tag, halotag}, month = {June}, posted-at = {2008-06-23 17:59:07}, priority = {2}, title = {HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein Analysis}, url = {http://dx.doi.org/10.1021/cb800025k}, year = {2008} }

TY - JOUR ID - citeulike:2870105 L3 - citeulike-article-id:2870105 TI - HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein Analysis JF - ACS Chem. Biol. AD - Promega Corporation, 2800 Woods Hollow Rd., Madison, Wisconsin 53711, !Promega Biosciences Incorporated, 277 Granada Dr., San Luis Obispo, California 93401, §Present address: National Renewable Energy Laboratory, 1617 Cole Blvd., Golden, Colorado 80401 N2 - We have designed a modular protein tagging system that allows different functionalities to be linked onto a single genetic fusion, either in solution, in living cells, or in chemically fixed cells. The protein tag (HaloTag) is a modified haloalkane dehalogenase designed to covalently bind to synthetic ligands (HaloTag ligands). The synthetic ligands comprise a chloroalkane linker attached to a variety of useful molecules, such as fluorescent dyes, affinity handles, or solid surfaces. Covalent bond formation between the protein tag and the chloroalkane linker is highly specific, occurs rapidly under physiological conditions, and is essentially irreversible. We demonstrate the utility of this system for cellular imaging and protein immobilization by analyzing multiple molecular processes associated with NF-ºB-mediated cellular physiology, including imaging of subcellular protein translocation and capture of proteinprotein and proteinDNA complexes. KW - fluorescence KW - fluorescent-tag KW - halotag AU - Los, Georgyi AU - Encell, Lance AU - Mcdougall, Mark AU - Hartzell, Danette AU - Karassina, Natasha AU - Zimprich, Chad AU - Wood, Monika AU - Learish, Randy AU - Ohana, Rachel AU - Urh, Marjeta AU - Simpson, Dan AU - Mendez, Jacqui AU - Zimmerman, Kris AU - Otto, Paul AU - Vidugiris, Gediminas AU - Zhu, Ji AU - Darzins, Aldis AU - Klaubert, Dieter AU - Bulleit, Robert AU - Wood, Keith PY - 2008/06/06/ UR - http://dx.doi.org/10.1021/cb800025k ER -