Expedeon’s Ni-NTA affinity resin is designed for simple, rapid His-tagged recombinant protein purification from a cell lysate under native or denaturing conditions. 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 which 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.
Benfits of Ni-NTA
- One step purification
- High capacity
- Purification under native or denaturing conditions
- Minimum metal leaching
Clarified E. coli lysate (10 ml) was purified on a 1ml Amintra NiNTA column (2,5 cm). Binding buffer: 50 mM Na2HPO4, 300 mM NaCl, 10 mM Imidazole, pH 8.0. Elution buffer: 50 mM Na2HPO4, 300 mM NaCl, 250 mM Imidazole, pH 8.0. Flow Rate: 1 ml /min.
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