Mee-Jung HAN, Meenhard HERLYN, Aron B. FISHER, David W. SPEICHER
언어
영어(ENG)
URL
https://www.earticle.net/Article/A115256
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원문정보
초록
영어
Current methods for quantitatively comparing complex proteomes such as two-dimensional gel electrophoresis (2-DE), fluorescent 2-D difference in gel electrophoresis (2-D DIGE), and liquid chromatography (LC)-mass spectrometry (MS) have limited resolution and dynamic ranges, and consequently can detect only a small portion of complex proteomes. Therefore, sample prefractionation is critical for more comprehensive coverage of the proteome and reliable detection of low abundant proteins. In this study, we report a 3-D DIGE method using DIGE labeling combined with microscale solution isoelectric focusing (MicroSol-IEF) fractionation and subsequent analysis on slightly overlapping narrow pH range 2-D gels. To illustrate its utility, this 3-D DIGE method was applied to analysis of human melanoma cell lines and mouse lung tissue extracts. This method has the advantage of much more extensive proteome coverage with detection of proteins over a wider concentration range compared with conventional 2-D DIGE alone. In addition, the use of fluorescent labeling prior to MicroSol- IEF avoids any complications resulting from slight run-to-run variations during MicroSol-IEF fractionations or the subsequent 2-D gel separations. Thus the combination of MicroSol-IEF and 2-D DIGE into a 3-D DIGE strategy produces a powerful method for more comprehensive and quantitative comparison of protein profiles of very complex proteomes.[This work was supported by National Institutes of Health Grants CA93372 and CA92725, as well as institutional grants to the Wistar Institute including an NCI Cancer Core Grant (CA10815), and grants from the Pennsylvania Department of Health]
키워드
MicroSol-IEF3-D DIGEComplex proteomes
저자
Mee-Jung HAN [ Dept. of Biomolecular and Chemical Engineering, Dongyang University ]
Meenhard HERLYN [ The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA. ]
Aron B. FISHER [ 2Institute for Environmental Medicine, University of Pennsylvania Medical Center, Philadelphia, PA 19104, USA. ]
David W. SPEICHER [ The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA. ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
1. 생물공학 분야의 발전을 위한 연구 협력
2. 생물공학의 실용화를 촉진시키기 위한 산학 협동
3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최
4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간
5. 생물공학 발전을 위한 정책 건의
6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동