通过分级沉淀和高效液相色谱法从鼠血清中快速、活性导向分离性别限制蛋白（Slp）

Rapid, activity-guided isolation of sex-limited protein (Slp) from mouse serum by fractionated precipitation and high-performance liquid chromatography.

作者信息

Van den Berg C W, Van Dijk H

机构信息

Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, UK.

出版信息

J Immunol Methods. 1994 Mar 10;169(2):251-6. doi: 10.1016/0022-1759(94)90269-0.

DOI:10.1016/0022-1759(94)90269-0

PMID:8133084

Abstract

We have recently demonstrated that sex-limited protein (Slp) plays a key role in an EDTA-resistant mouse complement activation pathway. A rapid procedure, utilizing classical chromatography methods on an FPLC system, was developed for the isolation of functionally active Slp. The method is based on the fractionated precipitation of serum by polyethylene glycol 6000, followed by heparin Sepharose Cl-6B affinity chromatography, Mono Q anion exchange and Superose 12 gel filtration. The isolation of Slp was monitored by a hemolytic assay. The procedure resulted in the purification of Slp, which by SDS-PAGE gave a single band of M(r) 2000,000 under non-reducing conditions, and under reducing conditions three bands corresponding to M(rs) of 105,000, 76,000 and 37,000.

摘要

我们最近证明，性别限制蛋白（Slp）在一种抗EDTA的小鼠补体激活途径中起关键作用。开发了一种利用FPLC系统上的经典色谱方法的快速程序，用于分离功能活性Slp。该方法基于用聚乙二醇6000分级沉淀血清，然后进行肝素琼脂糖凝胶CL-6B亲和色谱、Mono Q阴离子交换和Superose 12凝胶过滤。通过溶血试验监测Slp的分离。该程序导致Slp的纯化，通过SDS-PAGE在非还原条件下得到一条M(r)为2000,000的单带，在还原条件下得到三条分别对应于M(r)为105,000、76,000和37,000的带。