Purification of Growth-Promoting Peptides and Proteins,and of Histones,by High Pressure Silica gel Chromatography |
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Authors: | L R Pickart M M Thaler |
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Institution: | Department of Pediatrics and the Liver Center , University of California San Francisco , San Francisco, California, 94143 |
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Abstract: | A rapid method for the purification of histones and a variety of growth-promoting proteins and peptides by chromatography on silica gel has been developed. The isolation of the growth-promoting components of serum has been hampered by excessive losses associated with the use of water-based purification methods. The solubility of many growth-promoting serum proteins in acidic methanol-H2O solutions (eg. insulin, albumin, the somatomedins) provides a basis for purification on high-pressure silica gel columns, while peptides and histones can be purified in similar solvents. After column chromatography, the solvent is removed by flash-evaporation, or the protein may be precipitated directly from the solvent by neutralization of the pH and the addition of ethanol. The retention of biological activity (eg. somatomedin-C binding to insulin receptors and cell-growth stimulation) and recovery are excellent. |
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