title: Prestaining of glycoproteins in SDS-PAGE via 4H-[1]-Benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide with weak influence on protein mobility.
abstract: A new fluorescent prestaining method for gel-separated glycoproteins in 1D and 2D SDS-PAGE was developed by using 4H-[1]-Benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide (BH). The prestained gels were readily imaged after electrophoresis without any time-consuming steps needed for poststain. As low as 4-8 ng glycoproteins (transferrin, α1-acid glycoprotein) could be selectively detected, which is comparable to the most commonly used Pro-Q Emerald 488 glycoprotein stain. In addition, subsequent study of deglycosylation, glycoprotein affinity chromatography, and LC-MS/MS analysis were performed to confirm the specificity of the newly developed method. As a result, BH prestain provides a new choice for quick, sensitive, specific, economical, and MS compatible visualization of gel-separated glycoproteins.
authors.affiliation: School of Pharmaceutical Sciences, Key Laboratory of Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China; Wenzhou Undersun Biotechnology Co. Ltd, Wenzhou, Zhejiang, P. R. China.
authors.forename: Zhongxin
authors.initials: Z
authors.lastname: Zhu
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhou
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Qing
authors.initials: Q
authors.lastname: Yu
authors.forename: Xinliang
authors.initials: X
authors.lastname: Zhu
authors.forename: Chao
authors.initials: C
authors.lastname: Niu
authors.forename: Weitao
authors.initials: W
authors.lastname: Cong
authors.forename: Litai
authors.initials: L
authors.lastname: Jin
mesh_terms: D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D013876:Thiophenes / Q000737:chemistry*
chemical_list: D001798:Blood Proteins; D004396:Coloring Agents; D006023:Glycoproteins; D013876:Thiophenes; C016144:thiophene-2-carboxylic acid hydrazide
keywords: 4H-[1]-Benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide; Glycoprotein detection; Prestain