Synthesis and characterization of novel 4-(1-(4-(4-(4-aminophenyl)-1H-pyrazol-1-yl)-6-(4-(diethylamino)phenyl)-1,3,5-triazin-2-yl)-1H-pyrazol-4-yl)benzenamine fluorescent dye for protein binding

Novel 4-(1-(4-(4-(4-aminophenyl)-1H-pyrazol-1-yl)-6-(4-(diethylamino)phenyl)-1,3,5-triazin-2-yl)-1H-pyrazol-4-yl)benzenamine fluorescent dye was synthesized by multistep synthesis from cyanuric chloride and phenyl acetonitrile. It has absorption at 360 nm with single emission at 497 nm having fairly good quantum yield (0.379). The intermediates and the dye were characterized by FT-IR, 1H NMR, 13C NMR and Mass spectral analysis. Its utility as biocompatible fluorescent dye was explained by conjugation with bovine serum albumin. The method is based on direct fluorescence detection of fluorophore labelled protein before and after conjugation. Purified fluorescent conjugates were subsequently analyzed by fluorimetry. The analysis showed that the tested conjugation reaction yielded fluorescent conjugates of the dye through carbodiimide chemistry.