Cell Signaling HaloTag® Protein Purifi cation HaloTag® Protein Purifi cation System Product HaloTag® Protein Purifi cation System HaloTag® Protein Purifi cation System Sample Pack For Research Use Only. Not for Use in Diagnostic Procedures. Size Cat.# Price (Fr) 1 each G6280 966.00 1 each G6270 322.00 Description: The HaloTag® Protein Purifi cation System is designed to purify proteins fused to HaloTag® novel protein tag that enhances the expression and solubility of recombinant proteins. HaloTag® Technology enables the covalent, effi cient and specifi c capture of a protein of interest onto HaloLink™ Resin, thus overcoming the equilibrium-based limitations associated with affi nity tags (i.e., poor capture of proteins expressed at low levels and protein loss during washing of the purifi cation resin. The HaloTag® technology offers a quick and convenient way to test protein expression of HaloTag® fusion proteins as well as monitor the effi ciency of immobilization to HaloLink™ Resin by labeling with fl uorescent 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 purifi ed 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. Features: • Experience Superior Yield, Purity and Specifi c Activity of Soluble, Functional Proteins Compared to His-Tag, GST and MBP Affi nity Tags: Specifi c and covalent HaloTag® fusion protein capture and immobilization on HaloLink™ Resin. • Achieve Enhanced Target Protein Expression in Prokaryotic, Mammalian and Cell-Free Systems: Proteins are expressed as HaloTag® fusion proteins. • Purify Poorly Expressed Fusion Proteins: Rapid, specifi c and covalent capture of HaloTag® protein onto HaloLink™ Resin is a nonequilibrium process. • Effi ciently Recover Tag-Free Target Protein using TEV Protease Cleavage: Optimized TEV protease recognition site within the interconnecting polypeptide separating the HaloTag® protein and the fusion partner. HaloTag® protein remains immobilized on the resin due to covalent capture. • Save Time: One buffer compatible with downstream applications for all purifi cation steps. • Perform Easy In-Gel Detection and Quantifi cation of Protein Expression Levels with Fluorescent HaloTag® Ligands: Highly stable HaloTag® protein-ligand interaction permits boiling with SDS sample buffer followed by resolving on SDS-PAGE. Storage Conditions: Store the HaloLink™ Resin and HisLink™ Resin at 4°C. Do not freeze the resins. Store the TEV Protease at –20°C. Protocol HaloTag® Protein Purifi cation System Technical Manual Part# TM312 14 For complete and up-to-date product information visit: www.promega.com/catalog 245 Protein Purification and Interactions Pagina 248
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