title: Designing and Applying Proximity-Dependent Hybridization Chain Reaction.
abstract: Proximity-dependent hybridization chain reaction (proxHCR) is a novel technique for detection of protein interaction, post-translational modifications (PTMs), or protein expression. The method is based upon antibodies targeting the proteins of interest that are covalently conjugated to DNA oligonucleotides, which enables the induction of a hybridization chain reaction (HCR) to generate a fluorescent signal visible under a microscope. In contrast to the in situ proximity ligation assay (in situ PLA), which is another method that utilizes antibody-DNA conjugates to detect protein interactions, proxHCR does not require enzymatic steps. This makes proxHCR an inexpensive alternative to in situ PLA. Another potential advantage might be that proxHCR could more readily be adapted for use in automated staining procedures and in point-of-care devices, as all reagents can be stored at room temperature. This unit describes how the oligonucleotide system for proxHCR can be designed and a protocol for how to perform proxHCR is presented. © 2016 by John Wiley & Sons, Inc.
authors.affiliation: Max Planck Institute for Molecular Physiology, Department of Systemic Cell Biology, Dortmund, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Koos
authors.affiliation: Department of Immunology, Genetics and Pathology, Uppsala University, Science for Life Laboratory, Uppsala, Sweden.
authors.forename: Ola
authors.initials: O
authors.lastname: Söderberg
mesh_terms: D002460:Cell Line; D005456:Fluorescent Dyes; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis*; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes; D009841:Oligonucleotides; D011506:Proteins
keywords: hybridization chain reaction; post-translational modification; protein interaction; proxHCR; proximity ligation assay