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Electrophoresis. 2014 May;35(10):1439-47. doi: 10.1002/elps.201300595. Epub 2014 Apr 10.

Improved conditions for periodate/Schiff's base-based fluorescent staining of glycoproteins with dansylhydrazine in SDS-PAGE.

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1
Zhejiang Provincial Key Laboratory of Biopharmaceuticals, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China; Ningbo First Hospital, Ningbo, Zhejiang, P. R. China.

Abstract

An improved periodate/Schiff's base based fluorescent stain with dansylhydrazine (DH) for glycoproteins in 1D and 2D SDS-PAGE was described. Down to 4-8 ng of glycoproteins can be selectively detected within 2 h, which is approximately 16-fold higher than that of original protocol, but similar to that of Pro-Q Emerald 488 stain (Invitrogen, Carlsbad, USA). Furthermore, subsequent study of deglycosylation, glycoprotein affinity isolation, and LC-MS/MS analysis were performed to confirm the specificity of the improved method. As a result, improved DH stain may provide a new choice for selective, economic, MS compatible, and convenient visualization of gel-separated glycoproteins.

KEYWORDS:

Dansylhydrazine; Fluorescent stain; Glycoprotein detection

PMID:
24591039
DOI:
10.1002/elps.201300595
[Indexed for MEDLINE]
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