Purification of proteolipid protein and production of specific antiserum

Proteolipid protein, the major protein in CNS myelin, was purified using several chromatographic steps. No detergents were used. Chromatography in organic solvents on lipophilic Sephadex (LH-60) for delipidation, on ion exchange chromatography for protein separation, and again on lipophilic Sephadex (LH-20) for desalting, produced homogeneous preparations of proteolipid protein. Rabbit antibodies to proteolipid protein were produced after 2 injections of a total of 500 micrograms of protein. By immunotransfer and absorption experiments, it was found that the antiserum bound to proteolipid protein and a closely related protein, DM-20; it did not bind to basic protein or other myelin proteins.