RESUMO
We report the expedient microwave-assisted synthesis of thiophene based 4-sulfo-2,3,5,6,-tetrafluorophenyl esters whose molecular structure was engineered to achieve blue to red bright fluorescence. The reactivity toward monoclonal antibodies of the newly synthesized fluorophores was analyzed in comparison with that of the corresponding N-succinimidyl esters. Single-fluorophore and multiple-fluorophore labeled antibodies were easily prepared with both types of esters. Multiple-fluorophore labeling with blue and orange emitting fluorophores resulted in white fluorescent antibodies. Thiophene based fluorophores displayed unprecedented fluorescence stability in immunostaining experiments. First-principles TD-DFT theoretical calculations helped us to interpret the behavior of fluorescence emission in different environments.
Assuntos
Anticorpos Monoclonais , Corantes Fluorescentes/síntese química , Coloração e Rotulagem/métodos , Tiofenos/química , Anticorpos Monoclonais/química , Linhagem Celular , Humanos , Linfócitos , Micro-Ondas , Fixação de TecidosRESUMO
We report the synthesis of four oligothiophene-5-labeled deoxyuridines. These modified fluorescent nucleosides have been incorporated into oligodeoxynucleotides designed to be used as probes to discriminate, through changes in fluorescence emission, between hybridization with a perfectly complementary strand and that with a single nucleotide mismatch facing the modified uridines. Upon hybridization, remarkable differences (up to 47%) of the emitted light, depending on the uridine facing base, were observed.
Assuntos
Desoxiuridina/química , Corantes Fluorescentes/química , Testes Genéticos , Polimorfismo de Nucleotídeo Único/genética , Tiofenos/química , Pareamento Incorreto de Bases , Sequência de Bases , Desoxiuridina/análise , Fluorescência , Corantes Fluorescentes/análise , Corantes Fluorescentes/síntese química , Humanos , Oligonucleotídeos/genética , Compostos Organofosforados/química , Espectrometria de Fluorescência , Temperatura de Transição , Raios UltravioletaRESUMO
Oligomers of thiophene are widely studied compounds for their electronic and optoelectronic properties. Despite their strong fluorescence, their use as markers for biomolecules, especially for oligonucleotides (ONs), is still largely unexplored. Here, we describe the synthesis of a series of ON molecular beacons employing different oligothiophenes as fluorescent probes and discuss their fluorescence emissions in comparative experiments with and without dabcyl as a quencher, in their hairpin and linear conformations, and as duplexes after hybridization with a complementary target.
