Purification of human brain metallothionein by organic and reversed-phase high-performance liquid chromatography under acidic conditions

被引:7
作者
Cartel, NJ
机构
[1] Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ont. M5G 1X5
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 1996年 / 676卷 / 01期
关键词
metallothioneins;
D O I
10.1016/0378-4347(95)00409-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A simplified high-performance liquid chromatographic method for the detection of metallothioneins, notably metallothionein-III, has been developed. In order to purify metallothionein, differential acetone precipitation at 50% (v/v) and at 80% (v/v) was employed on a 20% normal human brain homogenate. The reconstituted pellet was injected into a C-18 microbore reversed-phase HPLC column, equilibrated with 0.1% trifluoroacetic acid, and developed at a flow-rate of 800 mu l/min with a linear gradient from 0% to 60% acetonitrile in 0.094% trifluoroacetic acid for 60 min. Western blots indicated that metallothioneins-I and II eluted at 16% acetonitrile and metallothionein-III eluted at 37% acetonitrile.
引用
收藏
页码:159 / 164
页数:6
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