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. 2016 Nov 29;4(4):36. doi: 10.3390/proteomes4040036

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© 2016 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).

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Proximity-dependent initiation of hybridization chain reaction (proxHCR). (a) Two proximity probes—i.e., antibodies conjugated to proximity hairpins (PH1 in green and PH2 in blue)—bind a protein complex (teal and ochre). Subsequently, an activator oligonucleotide is added (red line); (b) The activator breaks the secondary structure of PH1 open and binds to it, releasing its 3′-end; (c) The open PH1 can now invade PH2, thereby liberating its 3′-end. The latter can initiate the hybridization chain reaction (HCR); (d) The 3′ initiator sequence invades a fluorophore-labelled hairpin (teal line with a red star); (e) which then commences a chain reaction of invading and activating other such hairpins; (f) The reaction continues until depletion of the available fluorescent hairpins; (g) ProxHCR microscopy image: detection of interaction between E-cadherin and β-catenin in HaCaT cells. ProxHCR signal is shown in red and nuclei are stained in blue with Hoechst.

Proximity-dependent initiation of hybridization chain reaction (proxHCR). (a) Two proximity probes—i.e., antibodies conjugated to proximity hairpins (PH1 in green and PH2 in blue)—bind a protein complex (teal and ochre). Subsequently, an activator oligonucleotide is added (red line); (b) The activator breaks the secondary structure of PH1 open and binds to it, releasing its 3′-end; (c) The open PH1 can now invade PH2, thereby liberating its 3′-end. The latter can initiate the hybridization chain reaction (HCR); (d) The 3′ initiator sequence invades a fluorophore-labelled hairpin (teal line with a red star); (e) which then commences a chain reaction of invading and activating other such hairpins; (f) The reaction continues until depletion of the available fluorescent hairpins; (g) ProxHCR microscopy image: detection of interaction between E-cadherin and β-catenin in HaCaT cells. ProxHCR signal is shown in red and nuclei are stained in blue with Hoechst.