Principle

Protino® Ni-IDA products enable fast and convenient purification of recombinant polyhistidine-tagged proteins by immobilized metal ion affinity chromatography (IMAC). Protino® Ni-IDA is a dry silica-based resin precharged with Ni²⁺ ions. Binding of protein is based on the interaction between the polyhistidine tag of the recombinant protein and immobilized Ni²⁺ ions. The chelating group of Protino® Ni-IDA is based on IDA (iminodiacetic acid), which enables strong and efficient binding of target protein onto the IMAC matrix. In contrast to traditional IDA matrices, Protino® Ni-IDA shows an optimized, low density of IDA ligands, which is created by a special manufacturing process. This non-saturating surface concentration of IDA eliminates non-specific interactions of contaminating proteins with the adsorbent. As a result, Protino® Ni-IDA ensures higher target protein purity.

IDA is a threedentate chelator which occupies three of the six binding sites in the coordination sphere of the Ni²⁺ ion. The remaining three coordination sites are usually occupied by water molecules and can be exchanged with histidine residues of the recombinant protein.

Features

* Binding capacity will vary for each polyhistidin-tagged protein