DETECTION OF MYELIN BASIC-PROTEIN ISOFORMS BY ORGANIC CONCENTRATION

An effective technique was developed, which allowed rapid isolation of highly pure myelin basic protein (MBP) including its distinct isoform s. The procedure employs homogenization of central nervous system (CNS ) tissue in chloroform, which specifically extracts MBP. Subsequently, methanol was used to convert the protein susceptible to quantitative transfer into the acidic aqueous phase. MBP was purified from bovine, chicken, fish, human, guinea-pig, mouse, rabbit, rat, and swine brains . Analysis on SDS-PAGE and immunoblotting using polyclonal MBP-specifi c serum recognized proteins corresponding to the sizes of previously i dentified MBP isoforms of 21.5, 18.5, 17.2, and 14.2 kDa and three pre dicted isoforms of 20.2, 16.0, and 13 kDa. The MBP obtained was readil y soluble in water and possessed the capacity to induce experimental a utoimmune encephalomyelitis in susceptible mice. The protein was also suitable for use as a substrate for protein kinases. (C) 1997 Academic Press.