Promega Corporation

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.

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