Isolation and Purificationof Apolipoprotein A Ⅱ from Chicken Serum

摘要

ApoA Ⅱ in chicken serum was purified in common laboratory. HDL was isolated from pool serum by precipitating with Phosphotungstic acid-Mg. ApoA Ⅱ was obtained from HDL which was delipidated with Ethanol/Acetone (1:1, v/v) and permeated chromatography on DEAE Sepharose CL-6B column and Sephadex G-150 column, primarily identified by 15% Urea-SDS-PAGE. The purified apoA Ⅱ shows only one band on Urea-SDS-PAGE and the estimated molecular weight is 8,673u.The advantage of this method was very fussy by precipitating with phosphotungstic acid-Mg instead of ultracentrifugation, do not need special instruments. During the whole experimental procedures, we not only saved the precious time, but got a satisfied effect.