HistagNiNTA

HistagNiNTA refers to a specific type of affinity chromatography resin used in molecular biology for the purification of proteins that have been engineered with a polyhistidine tag. The resin is composed of a solid support matrix, typically agarose or a similar inert polymer, to which nitrilotriacetic acid (NTA) molecules are covalently attached. These NTA molecules are then chelated with nickel (Ni2+) ions. The polyhistidine tag, a sequence of six or more histidine amino acids, has a high affinity for these immobilized nickel ions. When a protein lysate containing a His-tagged protein is passed over the HistagNiNTA column, the His-tagged protein binds to the nickel ions on the resin, while other proteins in the lysate do not bind as strongly and can be washed away. The bound His-tagged protein can then be eluted from the column by using a solution containing imidazole, which competes with the histidine tag for binding to the nickel ions, or by lowering the pH, which disrupts the histidine-nickel interaction. This method provides a highly efficient and specific way to isolate and purify recombinant proteins from complex biological mixtures.