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Updated: May 22, 2026

Detection of Heterodimerization of Protein Isoforms Using an in Situ Proximity Ligation Assay

Detection of Heterodimerization of Protein Isoforms Using an in Situ Proximity Ligation Assay

Published on: October 20, 2018

Investigating Protein Interactions in Single Cells by Proximity Ligation Assay (PLA).

Zoe E Gillespie¹, Saloni Modi¹, Mariia Cherednychenko¹

¹Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada.

Methods in Molecular Biology (Clifton, N.J.)

|May 20, 2026

Summary

Proximity ligation assay (PLA) identifies interacting proteins and their location. This study optimizes PLA for nuclear proteins in mouse stem cells and blastocysts, providing a robust protocol.

Keywords:

Bastocyst Embryonic stem cells Immunolabeling Mouse Protein–protein interaction Proximity ligation assay Trophoblast stem cells

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Published on: September 2, 2019

Area of Science:

Cell Biology
Molecular Biology
Developmental Biology

Background:

Protein-protein interactions are crucial for cellular functions.
Understanding these interactions requires methods that reveal both proximity and subcellular localization.
Transcription factors and nuclear proteins play key roles in gene regulation and cellular processes.

Purpose of the Study:

To optimize the Proximity Ligation Assay (PLA) protocol for nuclear proteins.
To demonstrate the utility of PLA in various mouse cell types, including stem cells and blastocysts.
To establish a reliable positive control for nuclear protein interactions using PLA.

Main Methods:

Optimization of the Proximity Ligation Assay (PLA) protocol.
Application of PLA in mouse embryonic stem cells, trophoblast stem cells, and blastocysts.
Development and validation of a positive control for nuclear protein interactions.

Main Results:

An optimized PLA protocol for detecting nuclear protein interactions in mouse stem cells and blastocysts was established.
The protocol demonstrated robust detection of protein proximity and sub-cellular localization.
A reliable positive control for nuclear protein interactions was successfully implemented.

Conclusions:

Optimized PLA is a powerful tool for studying nuclear protein interactions in developmental contexts.
The protocol provides valuable insights into the localization and interaction of key regulatory proteins.
This method facilitates a deeper understanding of cellular processes governed by protein interactions in early development.