苯基-琼脂糖phenyl-sepharose疏水亲和层析.pdfVIP

GE Healthcare Instructions 71-7080-00 AE Hydrophobic interaction chromatography Phenyl Sepharose CL-4B TM Phenyl Sepharose CL-4B is a separation medium for hydrophobic interaction chromatography (HIC). Substances are separated on the basis of their different hydrophobicity. The phenyl group is covalently coupled to a cross-linked 4% agarose matrix by ether linkage, giving a hydrophobic medium with minimal leakage and no ionic properties. Table 1. Medium characteristics Ligand density: 40 μlmole/ml drained medium Available capacity*: HSA (Mr 68 000) 15-20 mg/ml drained medium ß-lactoglobulin (Mr 18 000) 3-5 mg/ml drained medium Bead structure: 4% cross-linked agarose Bead size range: 45–165 μm Mean bead size: 90 μm Max linear flow rate**: 150 cm/h at 25°C, HR 16/10 column, 5 cm bed height pH stability***. Long term: 3–12 Short term: 4–12 Chemical stability: Stable in all commonly used aqueous buffers. Physical stability: Negligible volume variation due to changes in pH or ionic strength. Autoclavable: 121 °C for 30 min. Storage: 20% ethanol or 0.1 M NaOH at 4 °C to 30 °C * The binding capacity was estimated in 0.01 M phospate buffer, pH 6.8, 1 M (NH ) SO 4 2 4 3 volumetric flow rate (cm ) ** Linear flow rate = 2 column cross-section