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 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
Specifications
Supporting matrix |
Covalently coupled to agarose resin |
Charged metal ion |
Ni2+ |
Bead size range |
45-165 ¥ìm |
Recommended working pH |
pH 2.0-12.0 |
Max binding capacity |
50 mg His-tagged recombinant protein/ml resin |
Linear Flow rate |
Up to 300 cm/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 |
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.
Chemical Compatibility