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. 2018 Nov;123(1):e58.
doi: 10.1002/cpim.58. Epub 2018 Sep 20.

Proximity Ligation Assay (PLA)

Muhammad S Alam  1
Affiliations

Proximity Ligation Assay (PLA)

Muhammad S Alam. Curr Protoc Immunol. 2018 Nov.

Abstract

Proximity ligation assay (PLA), also referred to as Duolink® PLA technology, permits detection of protein-protein interactions in situ (at distances <40 nm) at endogenous protein levels. It exploits specific antibodies identifying (either directly or indirectly) the two proteins of interest and utilizes specific DNA primers covalently linked to the antibodies. A hybridization step followed by DNA amplification with fluorescent probes permit visualization of spots of proximity by fluorescence microscopy. Since the development of PLA in 2002, it has been increasingly used to detect the interaction between two proteins with high sensitivity and specificity. It is a simple and sensitive technique to study protein-protein interaction in cells. © 2018 by John Wiley & Sons, Inc.

Keywords: Duolink®; antibody; in situ assays; protein-protein interaction.

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Figures

Figure-1
Figure-1. Representative diagram of the Duolink® proximity ligation assay (PLA)
Step by step representation of the PLA.
Figure-2
Figure-2. ZAP-70 and TCR zeta interaction in Jurkat cells
Confocal images of in situ PLA using rabbit anti-ZAP-70 and mouse anti-TCR zeta antibody in Jurkat cells that had been stimulated with anti-CD3/CD28 for 15 min.
See this image and copyright information in PMC

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