HaloTag® Technology for Protein Purification from Mammalian Cells

 

Get more protein with HaloTag®

It is all-too-often challenging to extract sufficient protein yields when expressing recombinant proteins in mammalian cells. Traditionally, the use of older fusion tag systems, like: FLAG® and His tags have been attempted, but often come up short in satisfaction. The FLAG® tag system can be frustrating given the resin’s low binding capacity, and the extremely high price. Alternatively, His-tagged proteins in mammalian lysates often results in contaminating endogenous His-rich proteins and metal (Ni) ion leakage that can cause problems with total recovery, purity and functionality of the target protein.

The HaloTag® platform can mitigate these common problems while maximizing protein yield, purity and cost savings.

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 HaloTEV Protease.
• Step 4: Recover POI without a tag.

Better recovery of protein compared with FLAG® and His₆ Tags

Two human kinases were purified from HEK293T cells with four commonly used fusion tags: HaloTag, FLAG , 3xFLAG and His₆ tag

Simple, cost-effective scalability

• Scalable protocols to generate mg-quantities of protein.
• Cost-effective resin for large-scale purification.

References

1. HaloTag-based purification of functional human kinases from mammalian cells.
2. Automethylation of CARM1 allows coupling of transcription and mRNA splicing.