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硝酸纤维素膜和凝胶上糖蛋白的鉴定

Identification of glycoproteins on nitrocellulose membranes and gels.

作者信息

Thornton D J, Carlstedt I, Sheehan J K

机构信息

Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, School of Biological Sciences, UK.

出版信息

Mol Biotechnol. 1996 Apr;5(2):171-6. doi: 10.1007/BF02789065.

Abstract

In this article we describe a procedure for the detection of glycoproteins on gels employing the periodic acid-Schiff's reagent. In addition, a number of staining protocols and direct binding ELISA, employing antibodies and lectins, are described for the identification and quantitation of glycoproteins after their immobilization by dot, slot, or Western blotting onto nitrocellulose membranes. We document, in detail, the conditions (i.e., the effect of solvent and detergents) for the immobilization of one specific family of O-linked glycoproteins, namely mucins. However, taking into account our suggestions, these procedures should be applicable to other types of glycoprotein.

摘要

在本文中,我们描述了一种使用高碘酸-席夫试剂检测凝胶上糖蛋白的方法。此外,还介绍了一些染色方案以及采用抗体和凝集素的直接结合酶联免疫吸附测定法,用于在通过斑点、狭缝或蛋白质印迹法将糖蛋白固定到硝酸纤维素膜上之后对其进行鉴定和定量。我们详细记录了固定一类特定的O-连接糖蛋白即粘蛋白的条件(即溶剂和去污剂的影响)。然而,考虑到我们的建议,这些方法应该适用于其他类型的糖蛋白。

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