Thermo Scientific™ Pierce Nickel Coated Plates provide a simple format to bind His-tagged fusion proteins for ELISA and other plate-based assays involving protein interactions of expressed recombinant proteins.
These Ni(2+) chelate-coated plates are ideal for analyzing polyhistidine-tagged fusion proteins by ELISA-based methods. Proteins that contain a succession of several histidine residues at the amino or carboxyl terminus have a strong binding affinity for metal. Bacterial lysates containing polyhistidine-tagged fusion proteins can be added directly to the plates without the need for blocking. The clear, white or black plates can be used with colorimetric, chemiluminescent or fluorescent detection methods, respectively.
Thermo Scientific Pierce Nickel Coated White 96-Well Plates provide a simple format to bind His-tagged fusion proteins for ELISA and other plate-based assays involving protein interactions of expressed recombinant proteins. These white plates can be used with chemiluminescent detection methods.
Features of Pierce Nickel Coated Plates include: • Ni2+ activated surface enables metal-chelate binding of His-tagged proteins • Detergents used to lyse cells don't inhibit binding to activated plates as they do with plain polystyrene • Better binding for sensitive assays compared to other commercially available nickel-activated plates • Detection limit: 1 ng of polyhistidine fusion protein • Binding capacity: approx. 9 pmol His-tagged protein (27 kDa) per well • Activation level: 200 μL
These Ni(2+) chelate-coated plates are ideal for analyzing polyhistidine-tagged fusion proteins by ELISA-based methods. Proteins that contain a succession of several histidine residues at the amino or carboxyl terminus have a strong binding affinity for metal. Bacterial lysates containing polyhistidine-tagged fusion proteins can be added directly to the plates without the need for blocking.
For Research Use Only. Not for use in diagnostic procedures.
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.