北京大学学报(自然科学版)

人胎肝金属硫蛋白的分离纯化及酶联免疫吸附检测

郑军恒;茹刚;茹炳根   

  1. 北京大学生命科学学院,北京,100871
  • 收稿日期:1997-12-31 出版日期:1999-03-20 发布日期:1999-03-20

Purification and Enzyme-Linked Immunosorbent Assay of Human Fetal Liver Metallothionein

ZHENG Junheng, RU Gang, RU Binggen   

  1. College of Life Sciences, Peking University, Beijing, 100871
  • Received:1997-12-31 Online:1999-03-20 Published:1999-03-20

摘要: 人胎肝经匀浆、离心、乙醇沉淀后,通过Sephadex G-75、DEAE-Sepharose Fast Flow柱层析分离,获得金属硫蛋白的两种亚型(MT-1和MT-2)。经氨基酸组成分析证明是均一的。MT-1为抗原免疫青紫兰兔制备了抗血清,以纯化的IgG建立了人MT的酶联免疫吸附检测(ELISA)方法,并给出了标准工作曲线。

关键词: 金属硫蛋白, 酶联免疫吸附检测, 分离纯化

Abstract: After human fetal liver was homogenized, centrifuged and precipitated with ethanol, two forms of metallothionein (MT-1, MT-2) were purified by gel filtration chromatography on Sephadex G-75, ion exchange chromatography on DEAE-Sepharose Fast Flow. Their purity was verified by amino acid composition analysis. Using human MT-1 as antigen, the antiserum was produced in rabbit. Then enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantitative analysis of human metallothionein, and the standard curves were supplied.

Key words: metallothionein, enzyme-linked immunosorbent assay (ELISA), purification and isolation

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