Large scale preparation and crystallization of neuron-specific enolase.
A simple method has been developed for the large scale purification of neuron-specific enolase [EC 4.2.1.11]. The method consists of ammonium sulfate fractionation of brain extract, and two subsequent column chromatography steps on DEAE Sephadex A-50. The chromatography was performed on a short (25 cm height) and thick (8.5 cm inside diameter) column unit that was specially devised for the large scale preparation. The purified enolase was crystallized in 0.05 M imidazole-HCl buffer containing 1.6 M ammonium sulfate (pH 6.39), with a yield of 0.9 g/kg of bovine brain tissue.
D000596:Amino Acids / Q000302:isolation & purification; D000645:Ammonium Sulfate; D000818:Animals; D001921:Brain / Q000201:enzymology; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002852:Chromatography, Ion Exchange; D003460:Crystallization; D009474:Neurons / Q000201:enzymology; D010751:Phosphopyruvate Hydratase / Q000302:isolation & purification*
D000596:Amino Acids; D010751:Phosphopyruvate Hydratase; D000645:Ammonium Sulfate