abstract
- The Proximity Ligation Assay (PLA) is a powerful method for visualizing protein-protein interactions with high specificity and sensitivity. By tethering DNA probes to antibodies directed against two target proteins, the PLA enables the detection of protein complexes within a 40-nm range. Upon colocalization of the target proteins, the DNA probes can anneal a connector oligonucleotide, initiating rolling circle amplification and generating a fluorescent signal detectable by microscopy. Here, we describe a detailed protocol for conducting a PLA in patient-derived tumor cells to visualize protein colocalization of two proteins in situ by fluorescence microscopy. With its ability to accurately capture protein-protein interactions and spatial organization within cells, the PLA is a valuable tool for insights into molecular mechanisms underlying brain tumor progression.