Darmanis Spyros, Kähler Anna, Spångberg Lena, Kamali-Moghaddam Masood, Landegren Ulf, Schallmeiner Edith
Department of Genetics and Pathology, Uppsala University, Sweden.
Biotechniques. 2007 Oct;43(4):443-4, 446, 448 passim. doi: 10.2144/000112551.
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.
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。