Choosing the right reagents is crucial when preparing to conduct affinity protein purification. After you’ve chosen a tag system to use, the next step is to choose an appropriate resin. This blog post will outline some of the differences between 4B and 6B agarose resin and how to choose the right resin for your needs.
Eluting your protein of interest is the final step in affinity chromatography before obtaining a pure, useful sample for further study. Common methods include elution using proteolytic elution, low pH elution, and denaturant elution. An alternative to these options is what’s known as Gentle Elution Buffer, or 3.6 M MgCl2 pH 6.6.
Low protein yield is a common issue in protein purification, especially when it comes to isolating large or complex structures. In lieu of using denaturants to elute protein from an affinity column, protease elution can be a welcome alternative to give tag-free protein.
This blog post about pH in the context of protein purification is the third in a series about optimizing conditions for protein purification. Although pH is most commonly seen as an issue in ion-exchange chromatography, it also plays a critical role in affinity tag protein purification.
When conducting affinity purification of a protein, there are many variables to consider in order to optimize purity and yield. One such condition to control is high salt loading, which decreases impurities in the final product.
