Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins

	Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins
	Liu, Fangjie 1,2; Ye, Mingliang 1; Wang, Chunli 1,2; Hu, Zhengyan 1,2; Zhang, Yi 1,2; Qin, Hongqiang 1,2; Cheng, Kai 1,2; Zou, Hanfa 1
刊名	analytical chemistry
	2013-08-06
卷号	85 期号:15 页码:7024-7028
ISSN号	0003-2700
通讯作者	邹汉法 ; 叶明亮
产权排序	1,1
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.
学科主题	物理化学
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
收录类别	SCI
语种	英语
WOS记录号	WOS:000323014000009
公开日期	2013-10-11
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/117567]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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.,...&Zou, Hanfa.(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.