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IMAC Sepharose 6 Fast Flow Media/HiTrap IMAC FF Columns/HiPrep IMAC FF 16/10 Column/HiScreen IMAC FF Column- For optimizing purification of histidine-tagged* proteins when Ni2+ is not the best choice of metal ion.
- Conveniently charge with your metal of choice.
- Ideal bead size for scaling up.
- High binding capacity.
- Prepacked HiTrap columns for convenient purification.
- Prepacked HiScreen columns for method optimization and process development.
- Prepacked HiPrep columns for easy scale-up.
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IMAC Sepharose 6 Fast Flow Media/HiTrap IMAC FF Columns/HiPrep IMAC FF 16/10 Column/HiScreen IMAC FF Column
Technical Information
* For licensing information, see Legal Info.
IMAC Sepharose™ 6 Fast Flow consists of 90-μm beads of highly cross-linked agarose to which a chelating group has been covalently coupled. This chelating group can be charged with suitable metal ions, specific for your target protein of interest.
The bead size of IMAC Sepharose™ 6 Fast Flow enables high flow properties, making the medium an ideal choice for scaling up purification at laboratory scale and purification of histidine-tagged proteins at process scale.
Low metal ion leakage means that the activity of the purified protein is retained and the risk of precipitation reduced, which results in increased purity, activity, and yield of the target protein. Leakage of metal ions into the eluted target protein pool from IMAC Sepharose™ 6 Fast Flow is generally low under normal conditions.
IMAC Sepharose™ 6 Fast Flow is highly stable and compatible with a wide range of common additives. This helps maintain biological activity and increase product yield, while at the same time greatly expanding the range of suitable operating conditions.
The medium is available prepacked in HiTrap™ HiScreen and HiPrep™ column formats for convenient purification of histidine-tagged proteins at laboratory scale. Initial purification using a 1-ml HiTrap™ IMAC FF column can be scaled up further with the 5-ml column. Purification can be scaled up even more using 20-ml HiPrep™ IMAC FF 16/10 columns. Prepacked HiScreen IMAC FF with a 10 cm bed height is the format of choice for method optimization and process development.
| TECHNICAL SPECIFICATIONS |
| Matrix | Highly cross-linked agarose, 6% |
| Average particle size | 90 μm |
| Binding capacity* | ~ 40 mg histidine-tagged protein/ml medium when charged with Ni2+ |
| Compatibility | Stable in all commonly used buffers |
| Chemical stability† | For one week at 40°C: 0.01 M HCl, 0.1 M NaOH
For 12 h: 1M NaOH, 70% acetic acid
30 min tested 30% 2-propanol
1 h tested 2% SDS |
| pH stability† | 3-12 (long term), 2-14 (short term) |
| Storage | 20% ethanol |
| Storage temperature | 4°C to 30°C |
| HiTrap™ prepacked columns |
| Column volume | 1 ml and 5 ml |
| Column dimensions | 0.7 × 2.5 cm (1 ml), 1.6 × 2.5 cm (5 ml) |
| Recommended flow rate‡ | 1 ml/min (1 ml), 5 ml/min (5 ml) |
| Max. pressure‡ | 3 bar (0.3 MPa, 42 psi) |
| HiScreen prepacked columns |
| Column volume | 4.7 ml |
| Column dimensions | 0.77 × 10 cm |
| Recommended flow rate‡ | 2.3 ml/min |
| Max. pressure over the packed | |
| bed during operation† | 1.5 bar (0.15 MPa, 22 psi) |
| Column hardware pressure limit | 5 bar (0.5 MPa, 73 psi) |
| HiPrep™ prepacked columns |
| Column volume | 20 ml |
| Column dimensions | 1.6 × 10 cm |
| Recommended flow rate‡ | 2-10 ml/min |
| Max. pressure over the packed bed during operation† | 1.5 bar (0.15 MPa, 22 psi) |
| Column hardware pressure limit | 5 bar (0.5 MPa, 73 psi) |
* Protein- and metal-ion dependent.
† Medium uncharged with metal ion.
‡ H2O at room temperature.
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