Nickel Agarose Resin is available in bulk or in prepacked columns. Ithas a binding capacity of >25 mg/ml with approximately 60 kDaprotein. In addition, it allows for purification of proteins undernative or denaturing conditions. GoldBio Nickel Agarose works very wellwith the His-Tag Buffer Set.
IDA cross-linked Agarose resin consists of iminodiacetic acid groups ligated by stable ether linkages via a spacer arm. IDA is a tridentate chelating agent, covalently coupled to cross-linked agarose beads. This resin is loaded with Ni2+. 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 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: Some discoloration of the Nickel IDA resin has been observed in reaction with low levels of reducing agents such as DTT. However, the resin remains fully functional even at concentrations as high as 5mM DTT.