Improved conditions for periodate/Schiff's base‐based fluorescent staining of glycoproteins with dansylhydrazine in SDS‐PAGE |
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| Authors: | Xuan Zhou Guo‐Ying Hong Bin‐Bin Huang Yuan‐Meng Duan Jia‐Yi Shen Mao‐Wei Ni Wei‐Tao Cong Li‐Tai Jin |
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| Affiliation: | 1. Zhejiang Provincial Key Laboratory of Biopharmaceuticals, Wenzhou Medical University, Wenzhou, Zhejiang, P. R. China;2. Ningbo First Hospital, Ningbo, Zhejiang, P. R. China;3. Wenzhou Undersun Biotechnology Co., Ltd, Wenzhou, Zhejiang, P. R. China |
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| 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. |
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| Keywords: | Dansylhydrazine Fluorescent stain Glycoprotein detection |
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