DICP OpenIR
Subject Area物理化学
Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins
Liu, Fangjie1,2; Ye, Mingliang1; Wang, Chunli1,2; Hu, Zhengyan1,2; Zhang, Yi1,2; Qin, Hongqiang1,2; Cheng, Kai1,2; Zou, Hanfa1; Zou HF(邹汉法); Ye ML(叶明亮)
Source PublicationANALYTICAL CHEMISTRY
2013-08-06
ISSN0003-2700
DOI10.1021/ac4017693
Volume85Issue:15Pages:7024-7028
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank1,1
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordMASS-SPECTROMETRY ; IDENTIFICATION ; PROTEOMICS ; DIGESTION ; MEMBRANES
AbstractTrypsin was immobilized on a variety of materials to improve digestion efficiency. However, because the immobilized trypsin will digest proteins during electrophoresis, direct immobilization of active trypsin in polyacrylamide gel will compromise the protein separation. To overcome this problem, here we report a novel polyacrylamide gel with switchable trypsin activity. It was prepared by copolymerization of the PEG-trypsin-aprotinin complex during the gel-casting step. Because the inhibitor aprotinin binds strongly with trypsin at alkaline pH, this novel gel does not display hydrolytic activity during electrophoresis. After electrophoresis, the activity of trypsin embedded in gel could be recovered by simply washing away the bound inhibitor at a low pH. It was demonstrated that this unique switchable activity design allowed high resolution of the complex protein mixture during electrophoresis and highly efficient digestion of the separated proteins in situ in the gel after electrophoresis.
Language英语
WOS IDWOS:000323014000009
Citation statistics
Cited Times:5[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/117567
Collection中国科学院大连化学物理研究所
Corresponding AuthorZou HF(邹汉法); Ye ML(叶明亮)
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Liu, Fangjie,Ye, Mingliang,Wang, Chunli,et al. Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins[J]. ANALYTICAL CHEMISTRY,2013,85(15):7024-7028.
APA Liu, Fangjie.,Ye, Mingliang.,Wang, Chunli.,Hu, Zhengyan.,Zhang, Yi.,...&叶明亮.(2013).Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins.ANALYTICAL CHEMISTRY,85(15),7024-7028.
MLA Liu, Fangjie,et al."Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins".ANALYTICAL CHEMISTRY 85.15(2013):7024-7028.
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