在尿素-十二烷基硫酸盐（SUDS）凝胶上分离的小肽的电免疫印迹法。应用于髓鞘碱性蛋白。

Electroimmunoblotting of small peptides separated on urea-dodecyl sulphate (SUDS) gels. Application to myelin basic protein.

作者信息

Sheng H Z, Martenson R E, Carnegie P R, Bernard C C

机构信息

Department of Psychology, La Trobe University, Bundoora, Vic., Australia.

出版信息

J Immunol Methods. 1988 Feb 24;107(1):13-22. doi: 10.1016/0022-1759(88)90003-8.

DOI:10.1016/0022-1759(88)90003-8

PMID:2449503

Abstract

A method for the electroimmunoblotting and immunodetection of peptides of less than 50 amino acid residues is described. Excellent resolution of a mixture of myelin basic protein (MBP) peptides was achieved by electrophoresis in a polyacrylamide stacking, urea-dodecyl sulphate minislab gel. Following electrophoresis, the peptides were transferred to various matrices and probed with monoclonal and polyclonal antibodies. Variables such as transfer time, membrane type, fixation and the amount of peptide loaded on the gel have been optimized as a consequence native and synthetic peptides can now be visualized in gels and immunodetected on immobilizing matrices. This procedure is particularly suited to the analysis and identification of small MBP fragments arising in various neuropathological conditions as well as for the rapid characterization of antigenic determinants recognized by monoclonal and polyclonal anti-MBP antibodies.

摘要

本文描述了一种用于对少于50个氨基酸残基的肽进行电免疫印迹和免疫检测的方法。通过在聚丙烯酰胺堆积、尿素-十二烷基硫酸钠微型平板凝胶中进行电泳，实现了髓鞘碱性蛋白（MBP）肽混合物的出色分离。电泳后，将肽转移到各种基质上，并用单克隆抗体和多克隆抗体进行检测。诸如转移时间、膜类型、固定以及凝胶上加载的肽量等变量已得到优化，因此现在可以在凝胶中观察到天然和合成肽，并在固定基质上进行免疫检测。该方法特别适用于分析和鉴定在各种神经病理条件下产生的小MBP片段，以及快速表征单克隆和多克隆抗MBP抗体识别的抗原决定簇。