Purification of myelin basic protein from bovine brain.
Myelin basic protein (MBP) is a commonly used substrate for in vitro determination of numerous protein kinase activities. Herein we describe a rapid method for isolating relatively large amounts of MBP from bovine brain with a purity greater than that currently available from commercial sources. Lipids were first extracted from the CNS tissue by homogenization in sec-butanol. Washes under neutral and mildly basic conditions were employed to remove neutral and acidic proteins from the defatted residue. MBP was subsequently extracted under acidic conditions and further purified by chromatography on CM Sephadex C-25. Potential contaminating enzyme activities were destroyed by heart treatment. This method typically yields a recovery of 1.0-1.5 mg MBP per gram of starting material with a purity of greater than 95%. The MBP prepared in this manner was suitable for determination of kinase activities by both solution and the "in gel" kinase assay systems.
Chevalier
D
D
Institut de Cardiologie de Montréal, Centre de Recherche, 5000 rue Bélanger est, Montréal, Québec, H1T 1C8, Canada.
Allen
B G
BG
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002847:Chromatography, Agarose; D004591:Electrophoresis, Polyacrylamide Gel; D004676:Myelin Basic Protein / Q000737:chemistry / Q000302:isolation & purification*
D004676:Myelin Basic Protein