Open AccessA Hybrid Detection Method Based on Peroxidase-mediated Signal Amplification and Click Chemistry for Highly Sensitive Background-free Immunofluorescent Staining
Author(s) -
Antonov Stanislav A.,
Novosadova Ekaterina V.,
Kobylansky Andrey G.,
Tarantul Vyacheslav Z.,
Grivennikov Igor A.
Publication year - 2019
Publication title -
journal of histochemistry and cytochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.971
H-Index - 124
eISSN - 1551-5044
pISSN - 0022-1554
DOI - 10.1369/0022155419864113
Subject(s) - click chemistry , immunostaining , staining , cycloaddition , chemistry , fluorescence , alkyne , azide , stain , microbiology and biotechnology , combinatorial chemistry , immunohistochemistry , biochemistry , catalysis , biology , organic chemistry , physics , quantum mechanics , immunology , genetics
The copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction is increasingly used for detection of various macromolecules and metabolites in biological samples. Here, we present a detailed analysis of the CuAAC reaction conditions in cells and tissue sections. Using the optimized CuAAC conditions, we have devised a highly sensitive immunostaining technique, based on the tyramide signal amplification/catalyzed reporter deposition (TSA/CARD) method with a novel alkyne tyramide substrate. The described method offers improved detection threshold compared to conventional immunofluorescent staining and produces significantly lower non-specific background than TSA/CARD with fluorescent tyramides.
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