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来自加州海兔单个已识别神经元的序列蛋白质分析。一种涉及聚丙烯酰胺梯度凝胶和等电聚焦的微电泳技术。

Sequential protein analysis from single identified neurons of Aplysia californica. A microelectrophoretic technique involving polyacrylamide gradient gels and isoelectric focusing.

作者信息

Rüchel R

出版信息

J Histochem Cytochem. 1976 Jul;24(7):773-91. doi: 10.1177/24.7.956643.

Abstract

Electrophoresis in polyacrylamide gradient gels and isoelectric focusing in polyacrylamide gels of capillary size are powerful tools for the analysis of molecular weight and charge properties of small protein samples. This is demonstrated using identified neurons from the abdominal ganglion of the sea hare Aplysia californica. Certain cell-specific peptides, which are considered to be neurosecretory, have been shown to be water soluble when ethylene glycol was employed as a mobilizing agent. Although the mode of action of ethylene glycol is not yet understood, this method may be of value for various extraction procedures. The application of a new staining method that is preferential for separations of sodium dodecyl sulfate-proteins yields information about the charge of water-insoluble proteins which has so far been inaccessible. Preliminary results gained by a small, two-dimensional mapping procedure as well as optical density separation patterns of two different nuclear protein fractionation from a single isolated nucleus outline further possibilties of the microgel techniques.

摘要

在聚丙烯酰胺梯度凝胶中进行电泳以及在毛细管尺寸的聚丙烯酰胺凝胶中进行等电聚焦,是分析小蛋白质样品分子量和电荷性质的有力工具。这一点通过使用海兔加州海兔腹神经节中已鉴定的神经元得到了证明。某些被认为是神经分泌性的细胞特异性肽,当使用乙二醇作为流动剂时,已被证明是水溶性的。尽管乙二醇的作用方式尚不清楚,但该方法可能对各种提取程序有价值。一种对十二烷基硫酸钠 - 蛋白质分离具有选择性的新染色方法的应用,产生了关于水不溶性蛋白质电荷的信息,而这些信息迄今为止是无法获得的。通过一个小型二维图谱程序以及从单个分离核中分离出的两种不同核蛋白分级分离的光密度分离模式获得的初步结果,概述了微凝胶技术的更多可能性。

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