COSMOSIL HIC
COSMOSIL 5HIC is designed for one step desalting and separation of proteins. Hydrophobic interaction chromatography (HIC) is an effective method for purification and separation of proteins (especially enzymes) based on differences in their surface hydrophobicity. Since this method does not use organic solvents like reversed phase chromatography, there is only a little loss in enzyme activity and the tertiary structure of proteins.
Material characteristics
Packing material | 5HIC |
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Silica gel | high purity porous spherical silica |
Average particle size | 5 µm |
Average pore size | approx. 300Å |
Specific surface area | approx. 150 m2/g |
Main interaction | hydrophobic interaction |
Application data
A buffer with high salt concentration, usually 1-2 mol/l of (NH4)2SO4, is used as an initial mobile phase for adsorption of samples to a weakly hydrophobic stationary phase. The elution is done with a decreasing salt gradient.
Condition | |
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Column size | 4.6 mm I.D. x 50 mm |
Mobile phase | A: 20 mmol/l phosphate buffer + 100 mmol/l Na2SO4 + 1.5 mol/l (NH4)2SO4 (pH6.0) B: 20 mmol/l phosphate buffer + 100 mmol/l Na2SO4 (pH6.7) B 0 → 100% /10 min linear gradient |
Flow rate | 1.0 ml/min |
Temperature | 30?oC |
Detection | UV 220 nm |
Sample |
|
Injection vol. | mixture 2.0 μl |
Separation of crude β-Glucosidase
Condition | |
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Column size | 4.6 mm I.D. x 50 mm |
Mobile phase | A: 20 mmol/l phosphate buffer + 100 mmol/l Na2SO4 + 1.5 mol/l (NH4)2SO4 (pH6.0) B: 20 mmol/l phosphate buffer + 100 mmol/l Na2SO4 (pH6.7) B 0 → 100% /10 min linear gradient |
Flow rate | 1.0 ml/min |
Temperature | 30?oC |
Detection | UV 220 nm |
Sample | Crude β-Glulosidase 10.0 mg/ml |
Injection vol. | 1.5 μl |
- Model: 04263-21
- Manufactured by: Nacalai Tesque