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Nat Commun. 2015 Jun 12;6:7294. doi: 10.1038/ncomms8294.

Proximity-dependent initiation of hybridization chain reaction.

Author information

1
Uppsala University, Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Biomedical center, Husargatan 3, Box 815, SE-75108 Uppsala, Sweden.
2
Department of Chemistry-BMC, Box 256, Uppsala University, SE-75123 Uppsala, Sweden.
3
School of Biological Sciences and School of Biomedical Sciences, University of Edinburgh, C H Waddington Building, Max Born Cresent, Kings Buildings, Edinburgh EH9 3BF, UK.
4
Institute of Pathology, Medical University of Graz, A-8036 Graz, Austria.

Abstract

Sensitive detection of protein interactions and post-translational modifications of native proteins is a challenge for research and diagnostic purposes. A method for this, which could be used in point-of-care devices and high-throughput screening, should be reliable, cost effective and robust. To achieve this, here we design a method (proxHCR) that combines the need for proximal binding with hybridization chain reaction (HCR) for signal amplification. When two oligonucleotide hairpins conjugated to antibodies bind in close proximity, they can be activated to reveal an initiator sequence. This starts a chain reaction of hybridization events between a pair of fluorophore-labelled oligonucleotide hairpins, generating a fluorescent product. In conclusion, we show the applicability of the proxHCR method for the detection of protein interactions and posttranslational modifications in microscopy and flow cytometry. As no enzymes are needed, proxHCR may be an inexpensive and robust alternative to proximity ligation assays.

PMID:
26065580
PMCID:
PMC4490387
DOI:
10.1038/ncomms8294
[Indexed for MEDLINE]
Free PMC Article

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