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从脑组织学切片直接免疫印迹到硝酸纤维素膜上:用神经垂体素蛋白进行评估

Direct immunoblotting from histological sections of brain onto nitrocellulose: evaluation with the protein neurophysin.

作者信息

Reisinger P W, Unger J W

机构信息

Department of Anatomy, University of Munich, Federal Republic of Germany.

出版信息

Electrophoresis. 1990 Aug;11(8):631-4. doi: 10.1002/elps.1150110809.

Abstract

A method for the isolation and localization of proteins and peptides from histological sections of rat and human brain by immunoblotting is described. For validation, the well-characterized protein neurophysin was electrophoretically transferred from formaldehyde-fixed or fresh tissue sections onto a nitrocellulose membrane. Neurophysin on the nitrocellulose membrane was detected by a specific antibody reaction. The antibody against neurophysin was visualized either by using secondary antibodies, conjugated with peroxidase or by protein A gold, followed by enhancement with silver. With this simple and fast method, neurophysin (or other proteins and peptides) can be identified on nitrocellulose membranes in areas that correspond to anatomically defined regions. Since the procedure combines the advantages of precise regional localization of polypeptides with the specificity of antibody-antigen reactions, the method may prove useful for rapid screening of the distribution of peptides or proteins in (brain) tissue.

摘要

本文描述了一种通过免疫印迹法从大鼠和人类脑组织切片中分离和定位蛋白质及肽的方法。为进行验证,将特征明确的蛋白质神经垂体素从甲醛固定或新鲜组织切片上通过电泳转移至硝酸纤维素膜上。硝酸纤维素膜上的神经垂体素通过特异性抗体反应进行检测。抗神经垂体素抗体可通过与过氧化物酶偶联的二抗或蛋白A金进行可视化,随后用银增强。通过这种简单快速的方法,可在与解剖学定义区域相对应的区域的硝酸纤维素膜上鉴定出神经垂体素(或其他蛋白质和肽)。由于该方法结合了多肽精确定位区域的优势与抗体-抗原反应的特异性,该方法可能对快速筛选(脑)组织中肽或蛋白质的分布有用。

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