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学科主题物理化学
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)
刊名ANALYTICAL CHEMISTRY
2013-08-06
ISSN0003-2700
DOI10.1021/ac4017693
85期:15页:7024-7028
收录类别SCI
文章类型Article
部门归属18
项目归属1809
产权排名1,1
WOS标题词Science & Technology ; Physical Sciences
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]MASS-SPECTROMETRY ; IDENTIFICATION ; PROTEOMICS ; DIGESTION ; MEMBRANES
英文摘要Trypsin 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.
语种英语
WOS记录号WOS:000323014000009
引用统计
被引频次:5[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/117567
专题中国科学院大连化学物理研究所
通讯作者Zou HF(邹汉法); Ye ML(叶明亮)
作者单位1.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
推荐引用方式
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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