For versatile magnetocapture assays using 6xHis-tagged biomolecules
Ni-NTA Magnetic Agarose Beads introduce greater flexibility into metal-chelate每based assay design. They bridge the gap between purification-scale procedures using Ni-NTA metal-chelate affinity chromatography resins and microplate-based assays using Ni-NTA HisSorb modules. The power of Ni-NTA combined with the convenience of magnetic bead technology allows magnetocapture assays to be performed on any scale desired. Ni-NTA Magnetic Agarose Beads are ideal for assays involving biomolecular interactions (see figure "Magnetocapture assays with Ni-NTA Magnetic Agarose Beads.") and can be used in individual tubes or in 96-well microplates.
Protocols using Ni-NTA Magnetic Agarose Beads can be automated on BioRobot systems. Four different ready-to-run protocols are available for flexible combination of 96-well microscale purification and assay setup. An overview of these purification and assay protocols and how they can be combined to suit different applications is shown in Figure "Protein purification and assay protocols can be combined to suit different applications.". More details are provided in the Ni-NTA Magnetic Agarose Beads Handbook that is supplied with Ni-NTA Magnetic Agarose Beads. For purification of larger amounts of protein, refer to the Ni-NTA Superflow 96 BioRobot Kit.
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| Magnetocapture assays with Ni-NTA Magnetic Agarose Beads. |
Ni-NTA Magnetic Agarose Beads are precharged with nickel and are ready to use. They can be used in very small volumes 〞 as little as 10 µl can be used to bind 3 µg protein 〞 in 96-well format. The beads are separated by the QIAGEN 96-Well Magnet Type A (see photo) which holds them on the sides of the wells while buffers are exchanged to wash or elute the 6xHis-tagged proteins. The beads are easily resuspended and can be used for assays without the need for prior protein purification. Ni-NTA Magnetic Agarose Beads protocols can be fully automated on the BioRobot 3000 and BioRobot 8000 . The 12-Tube Magnet is used for separating the beads in single tubes. The mild conditions used for elution of the 6xHis-tagged protein allow captured molecules to be eluted as complexes with the 6xHis-tagged interacting partner or eluted separately (see figure "Protein purification and assay protocols can be combined to suit different applications.").
 12-Tube Magnet |
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| Micrograph of Ni-NTA Magnetic Agarose Beads showing magnetic particles inside the beads. |
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| Protein purification and assay protocols can be combined to suit different applications. |
Ni-NTA Magnetic Agarose Beads can be conveniently employed in many high-throughput assay applications:
Procedures using Ni-NTA Magnetic Agarose Beads can be fully automated on the BioRobot 3000 and BioRobot 8000 providing:
The automated protocols include native and denaturing purification as well as interaction and immunoassay procedures (see figure "Protein purification and assay protocols can be combined to suit different applications."). The purification protocols are easily combined with interaction protocols giving the flexibility needed for a broad range of applications including diagnostics, drug screening, and immunology.
| Specifications | |
| Binding capacity per well | 3每15 µg (125每625 pmol of a 24 kDa protein) |
| Form | 3每15 µg (125每625 pmol of a 24 kDa protein) |
| Bead size | Average 50 µm (range: 20每70 µm) |
紱釬忒聊: Ni-NTA Magnetic Agarose Beads Handbook 12/01 (PDF version, 653 KB)