Simple and Specific Dual-Wavelength Excitable Dye Staining for Glycoprotein Detection in Polyacrylamide Gels and Its Application in Glycoproteomics
Joint Authors
Chiang, Yu-Hsuan
Li, Ding-Tzai
Lin, Tzu-Chun
Wu, Yu-Jen
Chen, Yu-Kuang H.
Hsu, Jue-Liang
Chen, Ching-Chuan
Shi, Fong-Ku
Chen, Kuan-Hung
Lu, Ya-Ting
Source
Issue
Vol. 2011, Issue 2011 (31 Dec. 2011), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2011-09-29
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Abstract EN
In this study, a commercially available fluorescent dye, Lissamine rhodamine B sulfonyl hydrazine (LRSH), was designed to specifically stain the glycoproteins in polyacrylamide gels.
Through the periodate/Schiff base mechanism, the fluorescent dye readily attaches to glycoproteins and the fluorescence can be simultaneously observed under either 305 nm or 532 nm excitation therefore, the dye-stained glycoproteins can be detected under a regular UV transilluminator or a more elegant laser-based gel scanner.
The specificity and detection limit were examined using a standard protein mixture in polyacrylamide gels in this study.
The application of this glycoprotein stain dye was further demonstrated using pregnancy urine samples.
The fluorescent spots were further digested in gel and their identities confirmed through LC-MS/MS analysis and database searching.
In addition, the N-glycosylation sites of LRSH-labeled uromodulin were readily mapped via in-gel PNGaseF deglycosylation and LC-MS/MS analysis, which indicated that this fluorescent dye labeling does not interfere with enzymatic deglycosylation.
Hence, the application of this simple and specific dual-wavelength excitable dye staining in current glycoproteome research is promising.
American Psychological Association (APA)
Chiang, Yu-Hsuan& Wu, Yu-Jen& Lu, Ya-Ting& Chen, Kuan-Hung& Lin, Tzu-Chun& Chen, Yu-Kuang H.…[et al.]. 2011. Simple and Specific Dual-Wavelength Excitable Dye Staining for Glycoprotein Detection in Polyacrylamide Gels and Its Application in Glycoproteomics. BioMed Research International،Vol. 2011, no. 2011, pp.1-8.
https://search.emarefa.net/detail/BIM-990281
Modern Language Association (MLA)
Chiang, Yu-Hsuan…[et al.]. Simple and Specific Dual-Wavelength Excitable Dye Staining for Glycoprotein Detection in Polyacrylamide Gels and Its Application in Glycoproteomics. BioMed Research International No. 2011 (2011), pp.1-8.
https://search.emarefa.net/detail/BIM-990281
American Medical Association (AMA)
Chiang, Yu-Hsuan& Wu, Yu-Jen& Lu, Ya-Ting& Chen, Kuan-Hung& Lin, Tzu-Chun& Chen, Yu-Kuang H.…[et al.]. Simple and Specific Dual-Wavelength Excitable Dye Staining for Glycoprotein Detection in Polyacrylamide Gels and Its Application in Glycoproteomics. BioMed Research International. 2011. Vol. 2011, no. 2011, pp.1-8.
https://search.emarefa.net/detail/BIM-990281
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-990281