Nickel NTA Agarose Beads
Nickel-NTA Agarose Resin has a binding capacity of ~50 mg/ml. In addition, it allows for purification of proteins under native or denaturing conditions. GoldBio Nickel Agarose works very well with the His-Tag Buffer Set.
NTA cross-linked Agarose resin consists of nitrilotriacetic acid groups ligated by stable ether linkages via a spacer arm. NTA is a tetravalent chelating agent, covalently coupled to cross-linked agarose beads, providing a higher specificity and lower ion leaching than IDA linked resins. NTA resins have also been shown to be more robust in the presence of higher concentrations of EDTA, but may require a higher imidazole concentration for protein elution. This resin is loaded with Ni 2+. The resulting, ready-to-use resin is ideal for rapid purifications of His-tagged proteins.
Affinity Chromatography is based on the interaction between certain superficial protein residues (histidines, cysteines and to a lesser extent tryptophans), with transition metal cations, forming chelates.
Nickel-NTA agarose resin is suitable for use with FPLC, but with pressures no more than 20 kPa. Too much pressure or too fast of a flow rate will result in diminished performance.
Storage/Handling: Store at 4°C. Do not freeze.
Note: GoldBio recommends using Nickel NTA resins in the presence of elevated levels of reducing agents in order to ensure optimal purification of your target protein. Nickel IDA resins may show some discoloration in reaction with low levels of reducing agents but are still functional at low levels of reducing agents (≤5mM DTT).
|Storage/Handling||store at 4°C. Do NOT freeze.|
GoldBio His-Tag Column Prep Protocol
Quick and Simple His-Tag Column Prep protocol including Pouring and Packing an Agarose Column