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Product introduction
Ni-6FF(IMAC) can be used for separation and purification with the interactions between Ni2+and some amino acids (mainly include histidine, cysteine, tryptophan) on the side chain of protein, and it is suitable for separation and purification of His-tagged protein and biological molecules which interact with Ni2+.
Advantages
1. Quick and easy (one-step purification).
2. Wide application, simple-operation. Suitable for gravity column and prepacked column.
3. Lower Ni2+ abscission probability and wider compatibility with reagents when compared with Ni-6FF(IDA).
Performance index
Matrix | Highly cross-linked with 6% agarose |
Particle size range | 45-165 µm |
Average particle size | 90 µm |
Binding capacity | 45 mg (His-tagged protein)/mL (media) |
pH stability* | 3-12 (long-term) 2-14 (short term) |
Chemical stability* | 0.01M HCl, 0.01M NaOH (1 week) 1MNaOH, 70% Ethanol (12 hours) 2% SDS (1 hour) 30% Isopropyl alcohol (30 minutes) |
Flow rate | 300-600 cm/h (0.3MPa, XK16/40. Column bed height:30 cm) |
Pressure | ≤ 0.3 MPa |
Storage buffer | 20% Ethanol |
Storage temperature | 4~8℃ |
Compatibility with common reagents
Buffer | 0.05M sodium phosphate, pH 7.4 0.1M Tris-HCl, pH 7.4 0.1M Tris-acetate, pH 7.4 0.1M HEPES, pH 7.4 0.1M MOPS, pH 7.4 0.1M sodium acetate, pH 4 |
Denaturant | 8M Urea 6M Gua-HCl |
Detergent | 2% Triton X-100 2% Tween 20 2% NP-40 2% Cholate 1% CHAPS |
Reductant* | 0.005M DTE 0.005M DTT 0.02M β-mercaptoethanol 0.005M TCEP 0.01M reduced glutathione |
Other additives | 0.5M Imidazole 20% Ethanol 50% Glycerol 0.1M Na2SO4 1.5M NaCl 0.001M EDTA** 0.06M Citrate |
* During the operation with Ni-6FF(IMAC), it is allowed to add low concentration of reductant, but it must be avoided of being immersed or stored for a long time with solution with reductant.
** During the operation with Ni-6FF(IMAC), it is allowed to add nominal concentration of metal ions chelating agent (e.g. 0.0001M EDTA) into samples of small volume, but it must be avoided of adding or loading a large volume of samples containing chelating agent into the purified solution.
Application examples
Example 1:
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