According to the different binding groups, Ni²⁺affinity chromatography columns can be divided into two types: Ni IDA agarose gel and Ni NTA agarose IDA and NTA Png gel. Ni²⁺has six chelating valences, among which Ni-IDA chelates trivalent and Ni-NTA chelates tetravalent. Therefore, the load of IDA is higher than that of NTA, but its weak binding force makes metal ions easy to be leached during the elution stage and closely combine with the target protein, resulting in low yield of isolated protein, impure product and metal ion pollution. The particle size of NTA is uniform and smaller, and the nickel chelate is more stable, which can withstand higher reducing agents, making the filler more stable, and nickel ions are not easy to fall off.

If you are pursuing high yield and do not require high protein purity, you can choose nickel IDA agarose gel; If the protein purity is required to be high, nickel NTA agarose gel is better.

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