Ni-NTA Superflow 96 BioRobot Kit
For automated, medium-scale purification
of 6xHis-tagged proteins
| Yield: |
Medium-scale, up to 300 µg
protein per well |
| Lysate
clearing: |
TurboFilter 96 plates |
| Purification
module: |
QIAfilter 96 plates loaded with Ni-NTA Superflow |
| Throughput: |
96 samples in parallel |
| Processing: |
BioRobot
systems |
| Starting
material: |
5 ml cultures in 24-well blocks or tubes |
The
Ni-NTA Superflow 96 BioRobot Kit provides automated purification of up to 300
µg recombinant protein from 96 samples in parallel. The procedure uses a
TurboFilter 96 plate to clear bacterial cell lysates by vacuum filtration on a
BioRobot 3000, 8000, or 9600 workstation. The cleared lysates flow directly into
a QIAfilter 96 plate, which is preloaded with Ni-NTA Superflow by the BioRobot
workstation. This unique 96-well metal-chelate affinity-chromatography module
strongly and selectively binds 6xHistagged proteins. After several efficient
wash steps, the purified proteins are eluted in 450 µl elution buffer.
Ready-to-run protocols are provided for purification under native or denaturing
conditions. For purification of larger amounts of 6xHis-tagged protein, larger
cultures and several wells can be used (one well per 5 ml culture).
Automated
Expression-Clone Screening
 |
Medium-scale purification for expression-clone screening and protein
characterization. A mixture of vector constructs for the expression of
proteins of 12, 15, and 28 kDa, and a 48/50 kDa heterodimer was
transformed into E.coli, plated on selective medium, and 96 colonies
were picked at random for inoculating 5 ml cultures. Expression of
6xHis-tagged proteins was induced with IPTG overnight. Cells were
pelleted and lysed and 6xHis-tagged proteins purified. 5 µl of
each elution fraction was loaded for SDS-PAGE analysis; proteins were
visualized by Coomassie staining. |
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Ni-NTA
Superflow 96 BioRobot Kit 01/01 (PDF version, 486 KB)
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| Ordering
Information |
| Product |
Contents |
Cat. No. |
| Ni-NTA
Superflow 96 BioRobot Kit (24) |
For 24 x 96
6xHis-tagged protein preps: 24 QIAfilter 96 Plates, 24
TurboFilter 96 Plates, 3 x 100 ml Ni-NTA Superflow |
969263 |
|