In the post-genome sequencing era, researchers are increasingly turning their attention to the proteins encoded within the genome. How are their synthesis, degradation and conformation regulated? Do they interact with other proteins or nucleic acids or lipids? Can these interactions be perturbed? How do changes in the coding sequence of the gene affect the proteins and their function? Like DNA microarrays, protein arrays fulfill a need for miniaturization and throughput, but immobilizing proteins in a way that preserves function and conformation is not a simple problem to solve.
We introduce the HaloLink™ Protein Array Systems, which combine innovative HaloTag® Technology, surface engineering and cell-free protein expression to help you solve some of the problems of creating custom protein arrays.
The HaloTag® protein is a mutated hydrolase that forms a covalent bond with HaloTag® ligands. Under physiological conditions binding is rapid and highly specific, yielding a complex that is stable even under stringent conditions. Using the HaloLink™ Protein Array Systems, HaloTag® fusion proteins are expressed in a cell-free expression system and then covalently captured on hydrogel-coated glass slides derivatized with HaloTag® Ligands. The fusion proteins are captured directly from the expression reaction mixture without prior purification. Using this approach, multiple fusion proteins can be rapidly synthesized and immobilized in parallel on the slide surface, and an entire experiment including protein expression, custom array formation and protein interaction analysis can be completed in less than eight hours.
This video demonstrates how simple it is to synthesize your HaloTag® protein fusion and create a custom microarray suitable for a variety of downstream applications.
Latest posts by Michele Arduengo (see all)
- Popular Papers from Promega Authors - January 21, 2020
- MSI Testing of Tumor Cells for Better Tailored Treatment - November 7, 2019
- qPCR: The Very Basics - August 2, 2019