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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

Applications
• Purification of polyhistidine-tagged proteins
• Batch binding, gravity-flow column chromatography, MPLC / FPLC™ applications
Characterization:

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)
• High capacity and high affinity
• Simply replace your current Ni-NTA products, no optimization or protocol change necessary
• Purification under native and denaturing conditions
• Suitable for small proteins, large protein complexes, proteins with low expression rates
– universal use
• Market-leading performance for an unbeatable price

Protino® Ni-NTA Agarose
• Aqueous suspension
• For batch binding, gravity flow, and FPLC™ – highest flexibility of applications
• Empty Protino® Columns for gravity flow available


Technical Data:

Protino® Ni-IDA shows an optimized, low density of IDA ligands, which is created by a special manufacturing process. This non-saturating surface concentration of IDA eliminates non-specific interactions of contaminating proteins with the adsorbent. As a result, Protino® Ni-IDA ensures higher target protein purity.

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.

Structure of IDA in complex with Ni2+
Protino® Ni-IDA
gravity flow column procedure

Application data

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

 


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