Recombinant protein purification through the use of polyhistidine (His) tags and immobilized metal ion affinity chromatography (IMAC) is a highly favored method due to its simplicity and applicability to many different target proteins. His tags themselves are advantageous due to their high affinity to metal ion resins, and their small size minimizes any interference with protein activity, often making it unnecessary to even remove them. However, purification of His-tagged proteins using IMAC requires appropriate conditions to ensure sufficient purity and yield, and problems may arise if sample preparation and chromatographic processes are not handled properly. Here are a few tips to help you tackle any challenges and optimize your recombinant protein purifications.
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