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Purification of proteins by fractional interfacial salting out on unsubstituted agarose gels
Authors:Friedrich von der Haar
Institution:Max-Planck-Institut für Experimentelle Medizin Abteilung Chemie D-3400 Göttingen, Hermann-Rein-Straße 3 Germany
Abstract:Proteins can be precipitated onto the surface of unsubstituted agarose at an ammonium sulfate concentration about 10% lower than needed for precipitation out of solution. The protein is fractionally redissolved by developing agarose columns with a linear, decreasing gradient of ammonium sulfate. The method is characterized by high reproducibility, good purification factors and high recovery of enzymatic activity. As an example the method is applied to the purification of aminoacyl-tRNA synthetases (E.C. 6.1.1.-) specific for phenylalanine, isoleucine and valine.
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