When I was characterizing proteins in graduate school, my life was filled with constructs, constructs, constructs. I made a variety of subclones to synthesize and isolate parts and pieces of the protein in vitro. I made clones and subclones to generate a panel of antibodies against different parts of the protein. Some of those antibodies ended up working best on Westerns; others performed better in immunocytochemistry experiments. There was no one tool or tag that could be used for every step in the characterization of the protein.
Halotag® fusion tag changes that.
The HaloTag® fusion tag spans both worlds of looking at proteins in isolation for studies of protein interactions and post-translational modifications to studying proteins in cells through real-time imaging or localization studies.
The key to the HaloTag® Technology versatility is the covalent bond that it forms with its synthetic chloroalkane ligand. The HaloTag® protein is an engineered derivative of a dehalogenase protein originally isolated from Rhodococcus species. The choloralkane ligand can be conjugated to any one of many specific functional groups, providing a host of interchangeable ligands with different functionality. One clone, one fusion tag, many research applications.
Because of the interchangeable ligands, you can directly correlate imaging results to biochemistry data. The covalent binding of tag to ligand, allows a gentle protein isolation protocol, giving you the ability to capture many protein interactions using simple protocols and controls. And, you can even “see” protein interactions in live cells using Bioluminescence Resonance Energy Transfer (BRET) with the HaloTag® fusion tag and NanoLuc® luciferase.
There are several ways to begin exploring the HaloTag® technology. You can search the Find My Gene™ database for experession-validated human ORF clones that have been created with the HaloTag® fusion tag already attached. Or you can request a free HaloTag® expression vector card . Learn more about the HaloTag® Technology Platform today by visiting our Web site or viewing the archived webinar about the HaloTag® Technology. To see how other researchers are using HaloTag® Technology, visit the citations database at promega.com.