The Proximity Ligation Assay(PLA) requires specific antibodies directed to the protines of interest. The assay utilizes the Duolink kit as described below.
DuolinkR In Situ reagents from OlinkR Bioscience enable detection, visualization and quantification of individual proteins, protein modifications and protein interactions in tissue and cell samples prepared for microscopy. The target is detected using one or two primary antibodies depending on the application. In the case that two primary antibodies are used, they must have been raised in different species. A pair of oligonucleotide labeled secondary antibodies (PLA probes) generates a signal only when the two PLA probes have bound in close proximity, either to the same primary antibody or two primary antibodies that have bound to the sample in close proximity. The signal from each detected pair of PLA probes is visualized as an individual fluorescent spot. These PLA signals can be quantified (counted) and assigned to a specific subcellular location based on microscopy images.