His-tag is the most commonly used protein fusion tag. Current available nickel purification systems are of limited use for purifying His-tagged proteins expressed in rabbit reticulocyte lysate because of the copurification of hemoglobin present in rabbit reticulocyte lysate. This often limits the downstream applications of purified proteins in fluorescence-based functional assays or protein:protein interaction studies and also decreases the amount of proteins purified.
The MagZ™ Protein Purification System, is a unique system designed for the purification of expressed His-tagged protein from rabbit reticulocyte lysate. This protein purification system has the advantage over nickel-based immobilized metal ion affinity chromatography (IMAC) because it eliminates 99.9% of hemoglobin contamination.
The three basic steps involved in His-tagged protein purification from TNT® Reticulocyte Lysate using the MagZ™ System are shown in the figure. The steps are optimized for efficient recovery of His-tagged proteins without copurification of hemoglobin. The magnetic nature of the MagZ™ Binding Particles allows purification to be performed in a single tube, simplifying and shortening the purification process to less than 30 minutes.
References citing use of the MagZ™ Protein Purification System
- Muik, M. et al. (2008) J. Biol. Chem. 283, 8014–22.