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Synthesis and spectroscopic characterisation of new ESIPT fluorescent protein probes

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Abstract

Three new benzazole isothiocyanate fluorescent dyes, 2-(4′-isothiocyanate-2′-hydroxyphenyl)benzoxazole, 2-(4′-isothiocyanate-2′-hydroxyphenyl)benzothiazole and 2-(4′-isothiocyanate-2′-hydroxyphenyl)benzimidazole were synthesised, purified until optical purity grade and characterised by spectroscopic techniques. UV/VIS and steady-state fluorescence were also applied to characterise the photophysical behaviour of the dyes. These dyes exhibit an intense fluorescence emission with a large Stokes shift, inherent to the class of benzazoles which relax by the excited state intramolecular proton transfer (ESIPT) mechanism. The dyes were studied for labeling bovine serum albumin (BSA), resulting conjugates BSA-dye with a remarkable photostability under UV/VIS radiation in relation to classical protein labels. The resulting conjugates presented fluorescence in the blue–green region. Direct fluorescence detection of protein-labeled with those dyes after polyacrylamide gel electrophoresis indicates their potential use as fluorescent probes for proteins.

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Authors and Affiliations

  1. Universidade Federal do Rio Grande do Sul - Instituto de Química, Laboratório de Novos Materiais Orgânicos, Av. Bento Gonçalves 9500., CP 15003, CEP 91501-970, Porto Alegre, RS, Brazil

    Fabiano Severo Rodembusch, Fernando Paulus Leusin & Valter Stefani

  2. Departamento de Ciência dos Alimentos, Laboratório de Bioquímica e Microbiologia Aplicada, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500, CEP 91501-970, Porto Alegre, RS, Brazil

    Luis Fernando da Costa Medina & Adriano Brandelli

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  1. Fabiano Severo Rodembusch
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  2. Fernando Paulus Leusin
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  3. Luis Fernando da Costa Medina
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  5. Valter Stefani
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Correspondence to Valter Stefani.

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Rodembusch, F.S., Leusin, F.P., da Costa Medina, L.F. et al. Synthesis and spectroscopic characterisation of new ESIPT fluorescent protein probes. Photochem Photobiol Sci 4, 254–259 (2005). https://doi.org/10.1039/b409233c

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