Description
Covalent, Oriented Attachment of HaloTag® Fusion Proteins
HaloLink™ Resin provides a method for covalent, oriented attachment of HaloTag® fusion proteins onto a solid surface. The resin consists of a HaloTag® ligand bound to Sepharose® beads that specifically and rapidly binds HaloTag® fusion proteins. HaloLink™ Resin has high binding capacity. Due to covalent linkage, HaloTag® fusion proteins cannot be eluted from the resin, allowing extensive washing to remove nonspecifically bound protein without the danger of eluting HaloTag® fusion proteins. The binding rate is very rapid and equivalent to biotin-streptavidin.
HaloLink™ Resin enables stringent washing, minimizing nonspecific background, without the risk of dissociation of bound HaloTag® proteins. The resin enhances the detection of protein:protein interactions and enables binding of proteins at low concentrations. One milliliter of settled resin binds >7mg of HaloTag® fusion proteins.
The HaloLink™ Resin can be used in a variety of applications including: detection and analysis of protein:protein interactions (in vivo and in vitro), detection of enzymatic activity of immobilized HaloTag® fusions and one-step purification of fusion protein in conjunction with proteolytic cleavage. A variety of vectors for the expression of HaloTag® fusion proteins in bacterial, mammalian or cell-free systems are available.
Note: Size listed indicates the amount of settled resin.
Detection of protein:protein interactions using HaloLink™ Resin. Panels A and B illustrate two approaches to characterizing protein:protein interactions. Panel C shows an analysis of protein:protein interaction.
