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The Thermo Scientific HisPur Ni-NTA Spin Purification Kit contains 0.2-ml HisPur Ni-NTA spin columns, as well as buffers required to purify polyhistidine-tagged proteins. HisPur Ni-NTA resin is a high-capacity, high-performance nickel-IMAC resin for routine affinity purification of His-tagged fusion proteins.
Features of HisPur Ni-NTA Resin:
• High capacity—bind up to 60 mg of 6xHis-tagged protein per milliliter of resin
• Versatile—purify proteins using native or denaturing conditions
• Compatible—use with Thermo Scientific Cell Lysis Reagents and a variety of buffer additives
• Cost-effective—reuse the same batch of resin at least five times
• Easy to use—pre-formulated buffers available for kit formats
• Flexible—available in multiple formats including bulk resin, spin columns, chromatography cartridges and 96-well filter plates
The specially prepared support consists of beaded agarose derivatized with the nitrilotriacetic acid (NTA) chelation moiety and loaded with divalent nickel ions (Ni2+). The immobilized metal affinity chromatography (IMAC) resin provides exceptional binding capacity and performance for recombinant His-tagged protein purification.
The expression and purification of recombinant proteins is central to protein regulation, structure and function studies. The majority of recombinant proteins are expressed as fusions with short affinity tags, the most popular being the polyhistidine (6xHis) tag. The method used to purify recombinant His-tagged proteins is immobilized metal affinity chromatography (IMAC), consisting of chelating resins charged with either nickel or cobalt ions that coordinate with the histidine side chains.
HisPur Ni-NTA Resin effectively purifies high levels of overexpressed His-tagged fusion proteins from bacterial lysates, such as those that are attained with Thermo Scientific B-PER Bacterial Protein Extraction Reagents. The resin performs well in batch-binding and spin-column procedures at a variety of scales. Performance equals or exceeds popular Ni-NTA resins from other suppliers. HisPur Ni-NTA Resin is a high-quality, stable and resilient affinity support. Tests confirm that no decrease in performance occurs after at least five repeated uses. These data indicated that the resin is highly resistant to structural degradation or nickel ion leaching during normal use.
HisPur Ni-NTA Resin and HisPur Cobalt Resin are alternative forms of IMAC, the former using nickel and the latter using cobalt as the chelated metal ion responsible for His-tag binding. Ni-NTA resins have been the most common IMAC resin choice for 6xHis-tag protein purifications because of the four metal-binding sites on the chelate, which enables high-protein binding and low-metal ion leaching. Cobalt binds less strongly and therefore more discriminantly, enabling greater purity but usually less yield in His-tagged protein purification.
• Resin Slurries—crosslinked 6% beaded agarose; 10 mL, 100 mL, 500 mL bottles
• Spin Columns—0.2 mL (microcentrifuge), 1 mL, 3 mL columns
• Purification Kits—complete kits in all three column sizes
• Chromatography Cartridges—learn more and see HisPur Chromatography Cartridge data
• 96-well Spin Plates
HisPur™ Ni-NTA Resin
HisPur™ Ni-NTA Spin Columns, 0.2 mL