Ni-NTA Spin Kit and Columns
For fast, small-scale purification of 6xHis-tagged proteins
| Binding capacity: | 150 µg/spin column (7.5 nmol @ ~20 kDa) |  |
| Support: | Macroporous silica | |
| Bead structure: | Spherical, silica particles | |
| Bead size: | 16¨C24 µm | |
| Form: | Dry matrix packed in spin columns, precharged with Ni2+ |
Ni-NTA silica combines Ni-NTA with a macroporous silica support material optimized to suppress nonspecific hydrophobic interactions. Ni-NTA Spin Kit and Columns provide Ni-NTA silica in a convenient microspin format for easy preparation of multiple samples in parallel. Each spin column can purify up to 150 µg of 6xHis-tagged protein from cellular lysates in just 15 minutes. Ni-NTA Spin Columns allow processing of up to 24 samples in as little as 30 minutes. They provide a simple method for functional screening of engineered proteins, selection of clones expressing full-length translation products, and comparison of expression levels (see Figure "Purification at Different Expression Levels"). Like all Ni-NTA matrices, Ni-NTA spin columns can be used for one-step protein purification under native or denaturing conditions. The Ni-NTA Spin Kit is a complete kit for spin purification of 6xHis-tagged proteins.
Purification at Different Expression Levels
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The 6xHis-tagged mouse DHFR was expressed at the indicated levels in E. coli, purified from 3 ml cultures using the Ni-NTA Spin Kit under denaturing conditions, and eluted in buffer at pH 5.9. Fractions were visualized by Coomassie staining after SDS-PAGE. 5 µl (of each 200 µl eluate) was loaded. 1: cell lysate; 2: flow-through 3: first eluate; 4: second eluate. |
²Ù×÷ÊÖ²á: Ni-NTA Spin Handbook (PDF version, 152 KB)