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HaloTag Technology: a multifunctional technology enabling protein purification, analysis of protein interactions, cellular imaging, and surface display for overall study of protein function

In the post-genomic era a major objective is to fully characterize the human proteome involving identification of proteins, determination of their localization, modifications, interactions, activities and ultimately their function. Essential to this ambition is development of new affinity and imaging tools allowing comprehensive analysis of protein function. Here we present a new technology based upon the use of a protein fusion tag, termed HaloTag, which allows for highly specific, oriented, and covalent immobilization on surfaces, and also protein labeling based on covalent binding with a variety of fluorescent ligands. Because of the multifunctional nature of HaloTag technology, analysis of many different aspects of protein function can be successfully achieved using a single construct.


Abstract: "HaloTag Technology for Live Cell Imaging"
This presentation will give an overview of HaloTag� technology for imaging, highlighting its inherent strengths as an imaging tool. Using this technology any protein of interest, expressed as a HaloTag� fusion can be specifically and efficiently labeled with a choice of spectrally distinct fluorescent tags. These small tags, called ligands, are comprised of two parts; a linker that covalently binds to the HaloTag� protein and a fluorescent moiety. Given that the ligands for imaging are of distinct wavelengths and include cell permeant and non-permeant options, this system affords great flexibility. Not only does it allow �color� choice for each experiment, but also the option of using two distinct ligands in pulse-chase fashion, allowing direct observation of protein turn-over or trafficking within live cells. In addition, the covalent bond that forms between HaloTag� protein and the ligand withstands denaturation such that reliable fixed cell and gel-based analyses are possible. Lastly, the chemistry involved in creating a ligand is quite simple and accommodates the creation of targeted dyes of any kind. This will be shown in the development and successful cellular targeting of a micro environmental sensor and a dye attuned to high resolution microscopy.


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