Why should I use ultra-pure water for washing Pierce Nickel Coated Plates?
The water used for washing of microplates as well as all assay reagents, must be of absolutely ultra-pure quality. This means that no metal ions should be present in the water, as this will bind to the His-tag and thereby decrease the binding of the fusion protein to the nickel-chelate complex.
Is it possible to elute the bound His-tagged fusion protein/peptide from the surface of Pierce Nickel Coated Plates?
Yes. For instance, use imidazole in concentrations >500 mM in Tris at pH 7.5 or a high concentration of EDTA.
With Pierce Nickel Coated Plates, can any reagent interfere with the binding between the nickel-chelate complex and the His-tagged fusion biomolecule?
Ionic detergents (e.g., SDS) will interfere with the binding as well as high concentrations of chelating reagents like EDTA, EGTA, and very strong electron donors like metal ions.
Can I use Pierce Nickel Coated Plates to measure the amount of bound His-tagged fusion protein/peptide?
To determine the approximate amount of bound His-tagged fusion protein/peptide, a standard curve of a previously purified preparation can be applied.