Open AccessApplication of BisANS fluorescent dye for developing a novel protein assay
Author(s) -
Zsolt Datki,
Zita Oláh,
Lilla Mácsai,
Magdolna Pákáski,
Bence Gálik,
Gabor Mihaly,
János Kálmán
Publication year - 2019
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0215863
Subject(s) - bicinchoninic acid assay , fluorescence , chemistry , chromatography , bradford protein assay , protein detection , chemical biology , peptide , chelation , protease , fluorophore , biochemistry , combinatorial chemistry , biophysics , nanotechnology , enzyme , biology , organic chemistry , materials science , physics , quantum mechanics
In many biology- and chemistry-related research fields and experiments the quantification of the peptide and/or protein concentration in samples are essential. Every research environment has unique requirements, e.g. metal ions, incubation times, photostability, pH, protease inhibitors, chelators, detergents, etc. A new protein assay may be adequate in different experiments beyond or instead of the well-known standard protocols (e.g. Qubit, Bradford or bicinchoninic acid) in related conceptions. Based on our previous studies, we developed a novel protein assay applying the 4,4′-Dianilino-1,1′-binaphthyl-5,5′-disulfonic acid dipotassium salt (BisANS) fluorescent dye. This molecule has several advantageous properties related to protein detection: good solubility in water, high photostability at adequate pH, quick interaction kinetics (within seconds) with proteins and no exclusionary sensitivity to the chelator, detergent and inhibitor ingredients. The protocol described in this work is highly sensitive in a large spectrum to detect protein (100-fold diluted samples) concentrations (from 0.28 up to more than 100 μg/mL). The BisANS protein assay is valid and applicable for quantification of the amount of protein in different biological and/or chemical samples.