Protino® Ni-IDA Resin
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Protino® Ni-IDA is a dry silica-based resin precharged with Ni2+ ions. Binding of protein is based on the interaction between the polyhistidine tag of the recombinant protein and immobilized Ni2+ ions.
Best choice for higher yields of His-tag proteins
|• Purification of polyhistidine-tagged proteins
• Batch binding, gravity-flow column chromatography, MPLC / FPLC™ applications
|Protino® Ni-NTA, the cost-effective NTA products for His-tag protein purification|
• Chelating group NTA (nitrilotriacetic acid), two protein binding sites (also see Technologies)
Protino® Ni-NTA Agarose
IDA is a threedentate chelator which occupies three of the six binding sites in the coordination sphere of the Ni2+ion. The remaining three coordination sites are usually occupied by water molecules and can be exchanged with histidine residues of the recombinant protein.
gravity flow column procedure
|Purification of polyhistidine-tagged GFPuv using Protino® Ni-IDA and Ni-IDA Agarose
Recombinant GFPuv was expressed in E. coli, lysed, loaded onto each gravity flow column, and eluted by a stepwise imidazole gradient. Eluted fractions were analyzed by SDS-PAGE. Pure polyhistidine-tagged protein can be eluted from Protino® Ni-IDA (left panel) at much lower imidazole concentrations than from Ni-IDA Agarose (right panel). In addition, Ni-IDA Agarose releases contaminating proteins from 10 mM to 100 mM imidazole.
M = Marker proteins, CL = Cleared lysate