Discussion
Purification of 6×His-tagged protein using Ni-NTA column is an example of ________
Answer: A
It is an example of affinity chromatography.
Nickel ions held on the column by nitrilotriacetic acid (NTA) bind the imidazole side chains of the six consecutive histidines of the tag. The tagged protein is retained by this specific interaction while other proteins wash through, and it is then eluted with imidazole or a low pH. This is immobilised metal affinity chromatography (IMAC), a form of affinity chromatography.
Separation does not rest on hydrophobic surface patches (hydrophobic-interaction chromatography), on net charge (ion-exchange chromatography) or on molecular size (size-exclusion chromatography).
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