Ni-NTA Affinity Resin - Amintra

Ni- NTA Affinity Resin - Amintra (ab270549) is designed for simple, rapid His-tagged recombinant protein purification from a cell lysate under native or denaturing conditions. Metal chelate affinity chromatography is a rapid one-step purification, which removes most contaminants and can achieve purities close to homogeneity. The rapid purification protocols provided in this handbook for affinity chromatography permit the recovery of high levels of pure recombinant protein in minutes. Recombinant proteins purified using Ni- NTA Affinity Resin - Amintra may be used in a wide range of structure and activity-based laboratory procedures.

Immobilized Metal Affinity Chromatography (IMAC) technology was introduced by Porath et al (1975). The matrix is attached to chelating groups that immobilize transition metal ions such as Ni²⁺, Co²⁺, Cu²⁺, Zn²⁺ (Porath and Olin, 1983; Porath, 1988; Sulkowski, 1989). Certain amino acids such as histidine, tryptophan, cysteine and tyrosine can act as electron donors on the surface of the protein and bind reversibly to the transition metal ion. In the vast majority of instances, 6x histidine tag is engineered at the N or C terminus of the protein (Kd-10-13 at pH 8.0).

Ni²⁺ is the most widely used metal ion as most IMAC tags seem to have very high affinity for immobilized Ni²⁺. The simplicity of IMAC technology is extremely attractive as it lends itself to the bind, wash and elute mode of operation if the appropriate buffer formulations are selected. IMAC can often be used with samples without any pre-treatment e.g. buffer exchange step. The use of metal chelate affinity is widespread for the selective adsorption of engineered recombinant proteins and has largely superseded non-affinity methods of chromatography for purifying recombinant proteins.

Characteristics of the resin:

This product is manufactured by Expedeon, an Abcam company and was previously called Amintra Ni-NTA Affinity Resin. Expedeon product codes ANN0010 (10 mL medium), ANN0025 (25 mL medium) and ANN0100 (100 mL) medium are the same as the current sizes: 10 mL, 25 mL and 100 mL respectively.