Pre-staining of glycoprotein in SDS-PAGE by the synthesis of a new hydrazide derivative

• Published in: Proteomics (Weinheim) Vol. 15; no. 21; pp. 3623 - 3628
• Main Authors: Zhou, Ayi, Zhou, Tieli, Yu, Dongdong, Shen, Yingjie, Shen, Jiayi, Zhu, Zhongxin, Jin, Litai, Zhang, Huajie, Wang, Yang
• Format: Journal Article
• Language: English
• Published: Germany Blackwell Publishing Ltd 01.11.2015; Wiley Subscription Services, Inc
• Subjects: Avidin - analysis; Chromatography, Liquid; Electrophoresis; Electrophoresis, Gel, Two-Dimensional - methods; Electrophoresis, Polyacrylamide Gel - methods; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - chemistry; Glycoprotein detection; Glycoproteins - analysis; Glycoproteomics; Humans; Hydrazines - chemical synthesis; Hydrazines - chemistry; Orosomucoid - analysis; Pre-stain; Pro-Q Emerald 300; Staining and Labeling - methods; Tandem Mass Spectrometry; Transferrin - analysis; UGF202; Electrophoresis; Pre-stain; Glycoproteomics; Pro-Q Emerald 300; Glycoprotein detection; UGF202
• DOI: 10.1002/pmic.201400567
• ISSN: 1615-9853

In this study, a new hydrazide derivative (UGF202) was synthesized and introduced as a highly sensitive and selective fluorescent probe to pre‐stain glycoproteins in 1D and 2D SDS‐PAGE. As low as 0.5–1 ng glycoproteins (transferrin, α1‐acid glycoprotein, avidin) could be selectively detected, which is comparable to that of Pro‐Q Emerald 300 stain, one of the most sensitive and commonly used glycoprotein staining kit. In addition, the specificity of the newly developed method was confirmed by the study of de‐glycosylation, glycoproteins affinity enrichment and LC‐MS/MS, respectively. According to the results, it is concluded that UGF202 pre‐stain can provide an alternative for the visualization of gel‐separated glycoproteins.