邻近连接分析（PLA）。

Proximity Ligation Assay (PLA).

机构信息

Laboratory of Immune Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

出版信息

Methods Mol Biol. 2022;2422:191-201. doi: 10.1007/978-1-0716-1948-3_13.

DOI:10.1007/978-1-0716-1948-3_13

Abstract

Proximity ligation assay (PLA), also referred to as Duolink PLA technology, permits detection of protein-protein interactions in situ (<40 nm distance) at endogenous protein levels. It exploits specific antibodies identifying (either directly or indirectly) the two proteins of interest and takes advantage of specific DNA primers covalently linked to the antibodies. A hybridization step followed by a PCR amplification with fluorescent probes then permits visualization of spots of proximity by fluorescence microscopy.

摘要

邻近连接分析（PLA），也称为 Duolink PLA 技术，允许在原位（<40nm 距离）检测内源性蛋白质水平的蛋白质-蛋白质相互作用。它利用特异性抗体识别（直接或间接）两个感兴趣的蛋白质，并利用特异性 DNA 引物共价连接到抗体上。随后进行杂交步骤和荧光探针的 PCR 扩增，然后通过荧光显微镜观察接近的斑点。