HomeFAQsWhat are to the advantages of performing purification under denaturing conditions?
FAQ: What are to the advantages of performing purification under denaturing conditions?
Deciding whether to purify a His-tagged protein under denaturing or native conditions depends on protein location and solubility, accessibility of the His-tag and whether or not the biological activity must be retained for downstream applications. Purification under denaturing conditions is most commonly employed when the tertiary structure of the native protein sequesters the His-tag, rendering purification under native conditions inefficient, or to solubilize inclusion bodies. Denaturing conditions can also mitigate the activity of proteolytic enzymes, reducing the risk of degradation of the target protein throughout the purification.