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CuAAC or click chemistry reaction

KN Karine Nozeret
AB Alix Boucharlat
FA Fabrice Agou
NB Nienke Buddelmeijer
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A mixture of 50 or 100 μM Azido-Cy5/FAM (10 mM stock solution in DMSO); 1 mM tris(2-carboxyethyl)phosphine hydrochloride (TCEP) (50 mM freshly prepared stock solution in ddH2O); 0.2 mM tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA) (2 mM stock in a mixture of tertbutanol and DMSO (4:1)); 1 mM CuSO4-5H2O (50 mM freshly prepared stock in ddH2O) was added to the 20 µL Lnt reaction solutions and samples were incubated at room temperature for 1 h. After incubation, an aliquot of each sample was diluted in loading buffer and heated at 100 °C for 5 min and loaded onto home-made Tris-Tricine SDS-PAGE gels for separation of peptides or processed for detection by fluorescence spectroscopy.

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