Optimized His-Tag Protein Purification from Cell-Free Expression Reactions

4453MAHis-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. Continue reading

How to Use the Flexi® Vectors (Part 1 of 2)

pFC7K (HQ) Flexi® VectorNow that we understand what the numbers, letters and abbreviations mean in the Flexi® Vector naming convention, how does one choose from among the over 40 different vectors? First I will detail the uses for the 21 vectors in the 1–12 range (plus outlier pF25) and address the remaining vectors in a second entry. Continue reading