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一种简便的肌醇磷脂微量分析方法

A Simple and Rapid Micro-analytical Method for Phosphoinositides

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【作者】 马克里刘彦崔肇春

【Author】 Ma Keli; Liu Yan; CuiZhaochun (Department of Biochbostry,Dalian Medical University, Dalian 116027,China).

【机构】 大连医科大学生物化学教研室

【摘要】 分析磷脂酰肌醇循环(PIcycle)的磷脂组分常采用双向薄层层析法.建立了一个简单快速的单向薄层层析分离肌醇磷脂方法.首先采用不同的有机溶剂体系分别提取非多磷酸肌醇磷脂和多磷酸肌醇磷脂,然后用不同的层析展开体系,对两部分磷脂进行单向薄层层析分离.采用无载体(32)P标记实验对该方法分离效果进行了观察.此法适用于同位素标记和非标记样品中肌醇磷脂组分的比较分析及多磷酸肌醇磷脂的提取、纯化和定量.

【Abstract】 Analysis of all of the PI cycle phospholipids is usually performed by twodimensional thin-layer chromatography. A simple and rapid method for analyzing the phosphoinositides by one-dimensional thinlayer chromatography has been developed.The non-polyphosphoinositides in cells were first extracted with chloroform: methanol:water (8: 4: 3), and then the polyphosphoinositides were extracted with chloroform:methanol: water: concentrated HCI (100:100: 50: 1). Each part of phospholipids were further separated by one-dimensional thinlayer chromatography with different developing systems, i. e., chloroform: methanol:acetic acid: formic acid: water (70: 30:12 : 4: 2) for non-polyphosphoinositide phospholipids and chloroform: methanol: water:NH4OH (40: 48: 10: 5) for polyphosphoinositides. The method can be used for comparative analysis of phosphoinositides in small samples, and also for the extraction,separation and quantitation of polyphosphoinositides, separation and quantitation of polyphosphoinositides in samples which are not labelled with isotope.

【基金】 国家自然科学基金
  • 【文献出处】 生物化学与生物物理进展 ,PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS , 编辑部邮箱 ,1996年01期
  • 【分类号】Q545.03
  • 【下载频次】64
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