By Amersham Biosciences
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Extra resources for Affinity Chromatography
Leakage of ligands from an affinity medium is always a possibility, especially if harsh elution conditions are used. The multi-point attachment of protein A to Sepharose results in very low leakage levels over a wide range of elution conditions. 33 Purification options Binding capacity Maximum operating flow Comments HiTrap Protein A HP Human IgG, > 20 mg/column Human IgG, > 100 mg/column 4 ml/min (1 ml column) 20 ml/min (5 ml column) Purification of IgG, fragments and sub-classes. Prepacked columns.
Interaction takes place with human colostral IgA as well as human myeloma IgA2 but not IgA1. Some human monoclonal IgMs and some IgMs from normal and macroglobulinaemic sera can bind to protein A. Leakage of ligands from an affinity medium is always a possibility, especially if harsh elution conditions are used. The multi-point attachment of protein A to Sepharose results in very low leakage levels over a wide range of elution conditions. 33 Purification options Binding capacity Maximum operating flow Comments HiTrap Protein A HP Human IgG, > 20 mg/column Human IgG, > 100 mg/column 4 ml/min (1 ml column) 20 ml/min (5 ml column) Purification of IgG, fragments and sub-classes.
The clone was a kind gift from Dr. C. Fuller and S. Brasher, Department of Biochemistry, University of Cambridge, UK. 4 Flow: Approx. 4 ml/min Equipment: Syringe Electrophoresis: SDS-PAGE, PhastSystem, PhastGel 10–15, 1 µl sample, silver staining Purification in 8 M urea Lane 1. Low Molecular Weight Calibration Kit (LMW) Lane 2. Starting material, cell extract, diluted 20-fold Lane 3. Flow-through, diluted 10-fold Lane 4. Wash Lane 5. Elution (first two ml) Lane 6. Elution (last two ml) Lane 7.