title: New method for covalent fluorescent biomolecule labeling with hemicyanine dye.
abstract: Fluorescent chromophore, alkylamino-(tetra-hydronaphthalenylidene)- benzothiazolium derivatives (HBTN dyes), are proposed as covalent labels for proteins via aliphatic amino groups. Spectral-luminescent properties of 3-methyl-2-{(E)-[7-(methylamino)-4,4a,5,6-tetra-hydronaphthalen-2(3H)-ylidene]methyl}-1,3-benzothiazol-3-ium chloride (HBTN, R=Me) and its predecessor, 2-[(E)-(7-methoxy-4,4a,5,6-tetrahydronaphthalen-2(3H)-ylidene)methyl]-3-methyl-1,3-benzothiazol-3-ium chloride (ABTN), are studied for free dyes and in the presence of DNA and BSA. Considerable spectral-luminescent changes accompany the transformation of ABTN into HBTN that allows monitoring conjugation reaction. In presence of DNA and BSA the HBTN increases its emission in 15 and 4 times respectively and becomes strongly fluorescent. The conditions for labeling are developed and a model conjugate of HBTN dye with BSA is synthesized. It was shown that using of HBTN dye as a fluorescent label allows detection by eye of about 3 mug/band of BSA on polyacrylamide gel upon UV-irradiation.
authors.affiliation: Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, 150 Zabolotnogo St., 03143 Kyiv, Ukraine.
authors.forename: Olexander M
authors.initials: OM
authors.lastname: Kostenko
authors.forename: Vladyslava B
authors.initials: VB
authors.lastname: Kovalska
authors.forename: Kateryna D
authors.initials: KD
authors.lastname: Volkova
authors.forename: Pavel
authors.initials: P
authors.lastname: Shaytanov
authors.forename: Igor O
authors.initials: IO
authors.lastname: Kocheshev
authors.forename: Yuriy L
authors.initials: YL
authors.lastname: Slominskiy
authors.forename: Irina V
authors.initials: IV
authors.lastname: Pisareva
authors.forename: Sergiy M
authors.initials: SM
authors.lastname: Yarmoluk
mesh_terms: D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D013058:Mass Spectrometry; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects; D013050:Spectrometry, Fluorescence / Q000379:methods*; D014466:Ultraviolet Rays
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins; D012710:Serum Albumin, Bovine; D004247:DNA