title: Design and synthesis of new fluorescent probe for rapid and highly sensitive detection of proteins via electrophoretic gel stain.
abstract: A new fluorescent molecular probe, 2,2'-(1E,1'E)-2,2'-(4-(dicyanomethylene)-4H-pyrane-2,6-diyl)bis(ethene-2,1-diyl)bis(sodium benzenesulfonate) salt (1), possessing the cyanopyranyl moieties and two benzene sulfonic acid groups was designed and synthesized to detect proteins in solution and for high-throughput SDS-PAGE. Compound 1 exhibited no fluorescence in the absence of proteins; however, it exhibited strong fluorescence on the addition of bovine serum albumin as a result of intramolecular charge transfer. Compared with the conventional protocols for in-gel protein staining, such as SYPRO Ruby and silver staining, 1 achieves higher sensitivity, even though it offers a simplified, higher throughput protocol. In fact, the total time required for protein staining was 60-90 min under optimum conditions much shorter than that required by the less-sensitive silver staining or SYPRO Ruby staining protocols. Moreover, 1 was successfully applied to protein identification by mass spectrometry via in-gel tryptic digestion, Western blotting, and native PAGE together with protein staining by 1, which is a modified protocol of blue native PAGE (BN-PAGE). Thus, 1 may facilitate high-sensitivity protein detection, and it may be widely applicable as a convenient tool in various scientific and medical fields.
authors.affiliation: National Institute of Advanced Industrial Science and Technology, c/o Katayanagi Advanced Research Laboratories, Tokyo University of Technology, Tokyo, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Takagi
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Chimuro
authors.forename: Atsushi
authors.initials: A
authors.lastname: Shinohara
authors.forename: Nao
authors.initials: N
authors.lastname: Sakaguchi
authors.forename: Atsunori
authors.initials: A
authors.lastname: Hiratsuka
authors.forename: Kenji
authors.initials: K
authors.lastname: Yokoyama
mesh_terms: D000818:Animals; D001557:Benzenesulfonates / Q000138:chemical synthesis / Q000737:chemistry; D015153:Blotting, Western; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D009942:Organometallic Compounds / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D016622:Silver Staining; D013050:Spectrometry, Fluorescence; D014357:Trypsin / Q000737:chemistry
chemical_list: D001557:Benzenesulfonates; D005456:Fluorescent Dyes; D009942:Organometallic Compounds; D010446:Peptide Fragments; D011506:Proteins; C515601:Sypro Ruby; D012710:Serum Albumin, Bovine; D014357:Trypsin