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Prestaining of glycoproteins in SDS ‐ PAGE via 4H‐[1]‐Benzopyrano[4,3‐b]thiophene‐2‐carboxylic acid hydrazide with weak influence on protein mobility
Author(s) -
Zhu Zhongxin,
Zhou Xuan,
Wang Yang,
Yu Qing,
Zhu Xinliang,
Niu Chao,
Cong Weitao,
Jin Litai
Publication year - 2014
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201400351
Subject(s) - glycoprotein , hydrazide , chemistry , chromatography , gel electrophoresis , biochemistry , carboxylic acid , transferrin , electrophoresis , organic chemistry
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.