Amintra® Ni-NTA Affinity Resin
Amintra® Ni-NTA Affinity Resin 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.
Amintra® Ni-NTA affinity resin offers excellent flow properties and high protein binding capacity: in excess of 50mg protein / ml.
NTA is a tetradentate chelator that occupies four of the six binding sites in the coordination sphere of the nickel ion. The other two coordination sites are usually occupied by water molecules and can interact with histidine residues of the recombinant protein. This binding minimizes metal leaching during purification.
- One step purification
- High capacity
- Purification under native or denaturing conditions
- Minimum metal leaching.
The rapid purification protocols for affinity chromatography permit the recovery of high levels of pure recombinant protein in minutes. Large numbers of samples can be processed at the same time. Recombinant proteins purified using Expedeon’s Amintra® Ni-NTA affinity resin may be used in a wide range of structure and activity based laboratory procedures.
- Supporting matrix: Covalently coupled to agarose resin
- Charged metal ion: Ni2+
- Bead size range: 45–165µm
- Recommended working pH: pH 2.0–12.0
- Maximum binding capacity: 50mg HIS-tagged recombinant protein / ml resin
- Linear Flow rate: Up to 300cm / h (5cm diameter column, pressure 1 bar)
- Optimum Flow Rate: 1–10 ml / min
- Maximum pressure: 0.1MPa (1 bar)
- Chemical stability: High
- Solubility in water: Insoluble.
Store the Amintra® Ni-NTA resin at 2–8˚C. Do not freeze or store the resin at room temperature.
Clarified E. coli lysate (10ml) was purified on a 1ml Amintra® Ni-NTA column (2,5 cm). Binding buffer: 50mM Na2HPO4, 300mM NaCl, 10mM Imidazole, pH 8.0. Elution buffer: 50mM Na2HPO4, 300mM NaCl, 250mM Imidazole, pH 8.0. Flow Rate: 1ml / min.