Human platelet monoamine oxidase (MAO B), a membrane bound enzyme was purified to homogeneity by DEAE-Sephacel column chromatography, chromatofocusing, and high performance liquid chromatography (HPLC). The crucial purification step was HPLC on a anion exchange column (SynChropak AX 300). The HPLC column was eluted initially with potassium phosphate buffer (100 mM, pH 7. 4) for 10 min at a flow rate of 1. 0 ml/min, followed by a gradient (0-1%) of octyl- beta -D-glucopyranoside (octylglucoside) in the same buffer for 10 min, and finally with buffered octylglucoside (1%) for 40 min. The extent of homogeneity and the subunit M//r (approximately 59,000) of MAO B were determined by sodium-dodecyl sulfate polyacrylamide gel electrophoresis followed by silver staining for proteins.
|Original language||English (US)|
|Number of pages||13|
|Journal||Journal of Liquid Chromatography|
|State||Published - 1983|
ASJC Scopus subject areas
- Molecular Medicine