Fractionation by salting out. (a) The salt of choice, usually ammonium sulfate, is added to a solution of macromolecules to a concentration just below the precipitation point of the protein of interest. (b) After centrifugation, the unwanted precipitated proteins (red spheres) are discarded and more salt is added to the supernatant to a concentration suffi cient to salt out the desired protein (green spheres). (c) After a second centrifugation, the protein is recovered as a precipitate, and the supernatant is discarded.
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