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Prestaining of glycoproteins in sodium dodecyl sulfate polyacrylamide gels by dansylhydrazine
Authors:Yuanmeng Duan  Binbin Huang  Guoying Hong  Ayi Zhou  Litai Jin
Institution:1. Zhejiang Provincial Key Laboratory of Biopharmaceuticals, Wenzhou Medical University, , Wenzhou, Zhejiang, P. R. China;2. Wenzhou Undersun Biotechnology Co., Ltd, , Wenzhou, Zhejiang, P. R. China;3. Department of histology and embryology, Institute of neuroscience, Wenzhou Medical University, , Wenzhou, Zhejiang, P. R. China
Abstract:A new fluorescent prestaining method for gel‐separated glycoproteins in 1D and 2D SDS‐PAGE was developed by using dansylhydrazine in this study. The prestained gels could be easily imaged after electrophoresis without any time‐consuming steps needed for poststains. As low as 4–8 ng glycoproteins (transferrin, α1‐acid glycoprotein) could be selectively detected, which is comparable to that of Pro‐Q Emerald 488, one of the most commonly used glycoprotein stain. In addition, a subsequent study of deglycosylation, glycoprotein affinity isolation, and LC‐MS/MS analysis was performed to confirm the specificity of the newly developed method.
Keywords:Dansylhydrazine  Electrophoresis  Glycoprotein detection  Prestain  Technology
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