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HaloCHIP™ System: Mapping Intracellular Protein:DNA
Interactions using HaloTag® Technology
Danette D. Hartzell, Jacqui Méndez, Katharine Hoffman, Doug Storts, Marjeta Urh and Keith Wood
Promega Corporation
The HaloCHIP™ System is a novel method that covalently captures intracellular protein:DNA complexes without using antibodies.
DNA binding proteins of interest are expressed in cells as HaloTag® fusion proteins, crosslinked to DNA, and then captured on
HaloLink™ Resin, which forms a highly specific, covalent interaction with the HaloTag® moiety on the fusion protein. Due to the complete
covalent linkage established between the HaloLink™ Resin and the crosslinked protein:DNA complexes, the resin can be stringently washed
to remove nonspecific DNA and protein more effectively than is possible by coimmunoprecipitation. The crosslinks are reversed to release
purified DNA fragments from the resin. By improving specificity and reducing background interference during the isolation of protein:DNA
complexes, this new method effectively increases the signal-to-noise ratio to permit detection of small changes in protein binding within
a genome.
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