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Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics

文献类型：期刊论文

作者Wu, Huifeng1; Ji, Chenglong1,2; Wei, Lei1,2; Zhao, Jianmin1
刊名PROTEOMICS
出版日期2013-11-01
卷号13期号:21页码:3205-3210
关键词2DE Animal proteomics Ecotoxicoproteomics Protein extraction Reproducibility
ISSN号1615-9853
产权排序[Wu, Huifeng; Ji, Chenglong; Wei, Lei; Zhao, Jianmin] Chinese Acad Sci, Yantai Inst Coastal Zone Res YIC, Key Lab Coastal Zone Environm Proc,YICCAS, Shandong Prov Key Lab Coastal Zone Environm Proc, Yantai 264003, Peoples R China; [Ji, Chenglong; Wei, Lei] Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
通讯作者Wu, HF (reprint author), Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China. hfwu@yic.ac.cn
中文摘要In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics.
英文摘要In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics.
学科主题Biochemical Research Methods ; Biochemistry & Molecular Biology
研究领域[WOS]Biochemistry & Molecular Biology
关键词[WOS]HIGH-FAT DIET ; MYTILUS-GALLOPROVINCIALIS ; PROTEOMIC ANALYSIS ; STRESS
收录类别SCI
资助信息NSFC [21237004]; Natural Science Foundation of Shandong Province [2013JQF01012]; Key Deployment Program of Chinese Academy of Sciences [KZZD-EW-14-03]
原文出处http://dx.doi.org/10.1002/pmic.201200421
语种英语
WOS记录号WOS:000330251700014
公开日期2014-07-08
源URL[http://ir.yic.ac.cn/handle/133337/6937]  
专题烟台海岸带研究所_中科院海岸带环境过程与生态修复重点实验室
烟台海岸带研究所_海岸带生物学与生物资源利用所重点实验室
作者单位1.Chinese Acad Sci, Yantai Inst Coastal Zone Res YIC, Key Lab Coastal Zone Environm Proc,YICCAS, Shandong Prov Key Lab Coastal Zone Environm Proc, Yantai 264003, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
推荐引用方式
GB/T 7714		 Wu, Huifeng,Ji, Chenglong,Wei, Lei,et al. Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics[J]. PROTEOMICS,2013,13(21):3205-3210.
APA Wu, Huifeng,Ji, Chenglong,Wei, Lei,&Zhao, Jianmin.(2013).Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics.PROTEOMICS,13(21),3205-3210.
MLA Wu, Huifeng,et al."Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics".PROTEOMICS 13.21(2013):3205-3210.

入库方式： OAI收割

来源：烟台海岸带研究所

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