Thermo Scientific HisPur Ni-NTA Magnetic Beads are high-capacity nickel-IMAC beads for affinity purification of His-tagged fusion proteins in manual or automated formats.
Features of HisPur Ni-NTA Magnetic Beads:
• High capacity—equivalent or higher binding capacity than Ni-NTA magnetic beads from other suppliers • Low nonspecific binding—the bead surface is pre-blocked and the protocol provides optimized buffers for purification • Fast—protocol is completed in 1 hour • Scalable—process microliter to milliliter sample volumes • Versatile—purify proteins using native or denaturing conditions • Reagent compatible—can be used with common cell lysis reagents and a variety of buffer additives • Multiple formats—protein coupling to the beads and downstream applications can be performed both manually and on an automated platform (e.g., Thermo Scientific KingFisher Instruments)
The blocked magnetic bead surface is derivatized with the nitrilotriacetic acid (NTA) chelation moiety and loaded with divalent nickel ions (Ni2+). The immobilized metal affinity chromatography (IMAC) beads provide high binding capacity with very low background. The HisPur Ni-NTA Magnetic Beads can be used both manually with a magnetic stand as well as with automated platforms such as the Thermo Scientific KingFisher Instruments for high-throughput needs.
HisPur Ni-NTA Magnetic 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). These tagged proteins are overexpressed in a number of different systems, most commonly in bacteria, and purified from cell lysates such as those prepared using B-PER Bacterial Protein Extraction Reagents. Purification of His-tagged proteins is achieved using a NTA chelate charged with nickel that coordinates with the histidine side chains. The NTA chelate contains four metal-binding sites which allow for low metal ion leaching and high binding capacity. The protocol for the HisPur Ni-NTA Magnetic Beads has been optimized to allow for high purity of the isolated His-tagged protein. Performance is equivalent to or better than Ni-NTA magnetic beads from other suppliers.