Thermo Scientific™ Pierce™ Ni-NTA Magnetic Agarose Beads provide a fast, convenient method for purification of His-tagged recombinant proteins. The beads are incubated with cell lysate containing His-tagged protein and then magnetically separated from the supernatant manually or through automation using an instrument such as the Thermo Scientific™ KingFisher™ Flex Magnetic Particle Processor. Nonspecifically bound protein can be washed away before dissociating bound His-tagged protein with elution buffer. Automated instruments are especially useful for higher throughput purification and screening of purification conditions.

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Pierce Ni-NTA Magnetic Agarose Beads consist of highly crosslinked agarose beads embedded with magnetite and a covalently attached tetradentate nitrilotriacetic acid (NTA) chelator charged with divalent nickel ions. The density of the ligand on the magnetic agarose bead results in a binding capacity similar to or better than traditional agarose resins with the added feature of magnetic handling. Magnetic agarose beads are a valuable tool for small-scale (∼1 mg) purification of multiple His-tagged proteins and for scouting expression and purification conditions to be used in larger-scale purifications with agarose chromatography supports.

Features of Ni-NTA Magnetic Agarose Beads:
• High capacity—sufficient for both routine and demanding purification procedures; binds >70 mg of 6xHis-tagged protein per mL of settled resin
• High-performance and reproducible beads—non-aggregating, magnetite (Fe₃O₄), superparamagnetic beads provide exceptional uniformity for both manual and automated HTS purification applications
• Low non-specific binding—optimized purification protocol results in better tagged-protein purification
• Versatile—purify proteins using native or denaturing conditions
• Compatible—use with Thermo Scientific Cell Lysis reagents and a variety of buffer additives

The high-performance, magnetite-containing, superparamagnetic magnetic agarose resin is validated and optimized for use with high-throughput magnetic platforms, such as the KingFisher 96 and KingFisher Flex magnetic particle processors, but the beads also enable premium performance for simple benchtop purification applications using an appropriate magnetic stand.

Ni-NTA Magnetic Agarose Beads are used for small-scale affinity purification as well as high-throughput screening of recombinant His-tagged proteins. The polyhistidine tag is the most popular affinity tag and typically consists of six consecutive histidine residues (6xHis). Tagged proteins are overexpressed in a number of different systems, most commonly in bacteria, and purified from cell lysates such as those prepared using Thermo Scientific B-PER Bacterial Protein Extraction reagents. Purification of His-tagged proteins is achieved using an NTA chelate charged with nickel that coordinates with the histidine side chains. The NTA chelate contains four metal-binding sites that allow for low metal ion leaching and high binding capacity. The protocol for the Ni-NTA Magnetic Agarose Beads has been optimized to allow for high purity of the isolated His-tagged protein.