title: In situ proximity ligation assay for microscopy and flow cytometry.
abstract: The ability to study proteins and protein interactions inside cells and tissues is important for elucidating how cells function in health and disease. The in situ proximity ligation assay (in situ PLA) presented here can be used to visualize proteins, protein-protein interactions, and post-translational modifications in cells and tissues. The method is based upon the use of antibodies that target the proteins involved in an interaction; hence, the method has the advantage that it can be used in clinical specimens, providing localized, quantifiable single molecule detection in single cells. This unit describes how in situ PLA can be used with fluorescence microscopy and flow cytometry to study proteins (obtaining high sensitivity and specificity of detection) and protein interactions. It also includes information on expected results and information on how to troubleshoot the assay.
authors: Department of Immunology, Genetics, and Pathology, Science for Life Laboratory, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.
authors: Karl-Johan
authors: KJ
authors: Leuchowius
authors: Department of Immunology, Genetics, and Pathology, Science for Life Laboratory, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.
authors: Irene
authors: I
authors: Weibrecht
authors: Department of Immunology, Genetics, and Pathology, Science for Life Laboratory, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.
authors: Ola
authors: O
authors: Söderberg
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001681:Biological Assay / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009841:Oligonucleotides / Q000276:immunology; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational
chemical_list: D000906:Antibodies; D009841:Oligonucleotides