HaloTag® Technology for Protein Purification from E. coli

 

HaloTag® Protein Purification System

• HaloTag is a highly-effective solubility tag
• Yields more active protein
• Improved protein recovery due to highly specific, covalent binding

Protocol Overview - Purified Protein Free of Tag

• Step 1: Immobilize HaloTag fused protein of interest (POI) onto HaloLink™ Resin.
• Step 2: Wash away unbound proteins
• Step 3: Release POI by TEV Protease (HQ tagged).
• Step 4: Remove TEV Protease by HisLink™ Resin.
• Step 5: Recover POI without a tag.

 

Data from a set of 23 difficult-to-express proteins shows that the HaloTag® Protein Purification System yields soluble protein more frequently that His, GST, or MBP affinity tags.

cPKA activity was determined using Kinase-Glo® assay after purification with multiple fusion tags, including HaloTag.