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微凝胶中纳克级复杂蛋白质混合物的等电聚焦(作者译)

[Isoelectric focusing of complex protein mixtures in the nanogram range in microgels (author's transl)].

作者信息

Bispink G, Neuhoff V

出版信息

Hoppe Seylers Z Physiol Chem. 1976 Jul;357(7):991-7.

PMID:11166
Abstract

A method is described for isolectric focusing of complex protein mixtures in 2, 5 or 10 mul capillaries. For one separation only 15- 50 ng of a protein mixture is needed. Isoelectric focusing is finished after 10 min, staining takes 20 min and destaining approximately 30 min. Using defined mixtures of Servalyt from different pH ranges, isoelectric focusing can be adapted to the protein sample to be fractionated. Protein peaks separated by isoelectric focusing can be electrophoretically eluted and for further analysis refractionated directly in a microgradient gel. The resolution power of microisoelectric focusing is as good as that of the wellknown macroprocedure, as is demonstrated by isoelectric focusing of the water soluble proteins from cerebellum and heart, of rat and human serum and of a human oncocytoma of the thyroid gland.

摘要

本文描述了一种在2微升、5微升或10微升毛细管中对复杂蛋白质混合物进行等电聚焦的方法。一次分离仅需15 - 50纳克蛋白质混合物。等电聚焦10分钟后完成,染色需20分钟,脱色约30分钟。使用来自不同pH范围的Servalyt特定混合物,等电聚焦可适应待分离的蛋白质样品。通过等电聚焦分离的蛋白质峰可进行电泳洗脱,并直接在微梯度凝胶中进一步分馏用于后续分析。微等电聚焦的分辨率与著名的宏观方法一样好,这已通过对大鼠和人类小脑及心脏的水溶性蛋白质、大鼠和人类血清以及人类甲状腺嗜酸性细胞瘤进行等电聚焦得到证明。

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