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用于灵活且模块化邻近连接分析的邻近探针自组装

Self-assembly of proximity probes for flexible and modular proximity ligation assays.

作者信息

Darmanis Spyros, Kähler Anna, Spångberg Lena, Kamali-Moghaddam Masood, Landegren Ulf, Schallmeiner Edith

机构信息

Department of Genetics and Pathology, Uppsala University, Sweden.

出版信息

Biotechniques. 2007 Oct;43(4):443-4, 446, 448 passim. doi: 10.2144/000112551.

Abstract

Proximity ligation assay (PLA) is a recently developed strategy for protein analysis in which antibody-based detection of a target protein via a DNA ligation reaction of oligonucleotides linked to the antibodies results in the formation of an amplifiable DNA strand suitable for analysis. Here we describe a faster and more cost-effective strategy to construct the antibody-based proximity ligation probes used in PLA that is based on the noncovalent interaction of biotinylated oligonucleotides with streptavidin followed by the interaction of this complex with biotinylated antibodies.

摘要

邻近连接分析(PLA)是一种最近开发的用于蛋白质分析的策略,其中通过与抗体连接的寡核苷酸的DNA连接反应基于抗体对靶蛋白进行检测,从而形成适合分析的可扩增DNA链。在此,我们描述了一种更快且更具成本效益的策略,用于构建PLA中使用的基于抗体的邻近连接探针,该策略基于生物素化寡核苷酸与链霉亲和素的非共价相互作用,随后该复合物与生物素化抗体相互作用。

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