By Nordic BioSite…
Ni-IDA columns from Nordic BioSite
Overview
Ni-IDA Columns form superior tools for routine purification of recombinant polyhistidine-tagged proteins, enabling purification to be performed under native and denaturating conditions.
Nordic BioSite can supply Ni-IDA columns from leading German manuafacturer MoBiTec that feature 1 g silica-based resin based on iminodiacetic acid (IDA). This enables strong and efficient binding of target protein onto the IMAC matrix to provide a fast and convenient route to purification of recombinant polyhistidine-tagged proteins by gravity flow.
Ni-IDA column features
MoBiTec Ni-IDA Columns from Nordic BioSite feature a form-stable silica matrix precharged with Ni2+ ions. This allows purification on the principle of Immobilized Metal Ion Affinity Chromatography (IMAC).
Poly-histidine tags are often used for affinity purification of poly-histidine-tagged recombinant proteins expressed in Escherichia coli, Bacillus megaterium, Bacillus subtilis, and other prokaryotic expression systems. A His tag is an amino acid motif in proteins that consists of multiple histidine (His) residues, mostly located at the N- or C-terminus of the protein. Using anti-poly-histidine antibodies is a prevalent technique for detection of successfully expressed His-tagged proteins.
Binding of proteins is based on the interaction between the polyhistidine tag of the recombinant protein and immobilized Ni2+ ions. The chelating group of the Ni-IDA resin is based on IDA (iminodiacetic acid), which enables strong and efficient binding of target protein onto the IMAC matrix.
IDA is a tridentate 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.
Ni-IDA advantages
In contrast with traditional IDA matrices, Ni-IDA is an optimized matrix with low density of IDA ligands. This non-saturating surface IDA concentration eliminates almost all non-specific interactions of contaminating host proteins with the adsorbent.
This allow’s Nordic BioSites’ Ni-IDA columns to provide higher target protein purity and faster purification of His-tagged proteins.
Summary
- Excellent tool for routine purification of recombinant polyhistidine-tagged proteins
- Starting from diverse expression systems, e.g., E. coli, yeast, insect, and mammalian cells
- Purification under native and denaturating conditions
- Maximal binding capacity: 90 mg protein per column
- Protein recovery rate > 80%
- Improved target specificity by optimized silica-based Ni2+-IDA matrix
- Imidazol-free loading and washing buffer
- Columns are long-term storable when kept dry
Resources
Click on Ni-IDA columns from Nordic BioSite for more information.
Click on Nordic Biosite to contact the company directly.
Supplier Information
Supplier: Nordic Biosite
Address: Propellervägen 4A, 183 62 Täby, Sweden
Tel: +46 (0)8 544 433 40
Fax: +46 (0)8 756 94 90
Website: www.nordicbiosite.com