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.