邻近连接试验
寡核苷酸
原位
DNA
荧光
共轭体系
原位杂交
化学
结扎
生物
分子生物学
细胞生物学
生物物理学
生物化学
生物素化
信使核糖核酸
基因
物理
受体
有机化学
量子力学
聚合物
作者
Arianna Bellucci,Chiara Fiorentini,Michela Zaltieri,Cristina Missale,PierFranco Spano
标识
DOI:10.1007/978-1-4939-0944-5_27
摘要
The proximity ligation assay (PLA) is a sensitive and specific technique to visualize proteins, their posttranslational modifications and activation state as well as protein–protein interactions. The assay is based on the employment of proximity probes, composed by oligonucleotide-conjugated antibodies, to recognize a couple of specific targets. The binding of probes in close proximity allows for their hybridization by connector oligonucleotides, that can form a circular DNA strand. These DNA circles can then be amplified by polymerase chain reaction. Finally, the conjugation of fluorescence-labelled oligonucleotides with the amplification product allows for the localized detection of individual or interacting proteins in cells and tissues. Here, we describe the use of “in situ” PLA to visualize the localization of protein–protein interactions in intact tissues.
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