Work

Technical Considerations for Detecting Protein–Protein Interactions Using Proximity Ligation Assay

Year: 2025

Type: article

Abstract: Proximity ligation assay has been widely used to detect protein-protein interaction in cells and tissues. While with great sensitivity, its specificity was often neglected. Here, we report the existence of varying levels of false positives observed with this assay, most likely due to its high sensitivity. We also provide suggestions to minimize false positives for more accurate detection of protein-protein interactions, especially for membrane proteins. These suggestions include co-staining target proteins, using various negative controls and suitable antibodies, avoiding detergents if possible, and validating interactions with complementary methods.

Source: Journal of Proteome Research

DOI: 10.1021/acs.jproteome.4c00855

Authors: Zach Tower, Hao Chang

Institutions: University of Wisconsin–Madison, William S. Middleton Memorial Veterans Hospital

Language: English

Topic: Biotin and Related Studies

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: green

Funders: U.S. Department of Veterans Affairs, University of Wisconsin-Madison, National Institutes of Health, National Institute of General Medical Sciences