HaloTag Protein Purification System
The HaloTag Protein Purification System is designed to purify proteins fused to the HaloTag novel protein tag that enhances the expression and solubility of recombinant proteins. HaloTag Technology enables the covalent, efficient and specific capture of a protein of interest onto HaloLink Resin, thus overcoming the equilibrium-based limitations associated with affinity tags (i.e., poor capture of proteins expressed at low levels and protein loss during washing of the purification resin. The HaloTag technology offers a quick and convenient way to test protein expression of HaloTag fusion proteins as well as monitor the efficiency of immobilization to HaloLink Resin by labeling with fluorescent HaloTag TMR Ligand followed by SDS-PAGE analysis. Guidelines can be found in the HaloLink Resin Technical Manual #TM250, the HaloLink Protein Array Technical Manual #TM310 and the HaloCHIP System Technical Manual #TM075. Outline of Procedure: The HaloTag protein, a 34kDa mutated hydrolase, covalently attaches to HaloLink Resin via an immobilized chloroalkane ligand. TEV Protease cleaves the target protein from the HaloLink Resin. The TEV Protease, which has an N-terminal (HQ) tag, is removed from the protein of interest using HisLink Resin, and the purified protein of interest is recovered. The appropriate vector that encodes the HaloTag protein and expresses protein optimally in E. coli is pFN18A HaloTag T7 Flexi Vector (G2751) or pFN18K HaloTag T7 Flexi Vector (G2681). These vectors can be purchased separately.