Fluorescent staining of glycoproteins in sodium dodecyl sulfate polyacrylamide gels by 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide.
A fluorescent detection method for glycoproteins in SDS-PAGE by using 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide (BH) was developed in this study. As low as 4-8 ng glycoproteins (transferrin, α1-acid glycoprotein) could be specifically detected by the BH staining method, which is twofold more sensitive than that of the most commonly used Pro-Q Emerald 488 glycoprotein stain. Furthermore, the specificity of the newly developed stain for glycoproteins was demonstrated by 1-D and 2-D SDS-PAGE, deglycosylation, glycoprotein affinity enrichment and LC-MS/MS, respectively. According to the results, it is concluded that BH stain may provide new choices for convenient, sensitive, specific and economic visualization of gel-separated glycoproteins.
School of Pharmaceutical Sciences, Zhejiang Provincial Key Laboratory of Biotechnology Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, Zhejiang, China. jin_litai@126.com cwt97126@126.com.
Zhongxin
Z
Zhu
Xuan
X
Zhou
Yang
Y
Wang
Lisha
L
Chi
Dandan
D
Ruan
Yuanhu
Y
Xuan
Weitao
W
Cong
Litai
L
Jin
D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D053719:Tandem Mass Spectrometry; D013876:Thiophenes / Q000737:chemistry*
D005456:Fluorescent Dyes; D006023:Glycoproteins; D013876:Thiophenes; C016144:thiophene-2-carboxylic acid hydrazide