A new proteomics sample processing method of intracellular insoluble aggregates
| Project/Area Number |
15K14427
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Genome biology
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| Research Institution | 防衛大学校(総合教育学群、人文社会科学群、応用科学群、電気情報学群及びシステム工学群) |
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Principal Investigator |
Ichimura Tohru 防衛大学校(総合教育学群、人文社会科学群、応用科学群、電気情報学群及びシステム工学群), 応用科学群, 教授 (50213012)
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| Co-Investigator(Kenkyū-buntansha) |
竹清 貴浩 防衛大学校(総合教育学群、人文社会科学群、応用科学群、電気情報学群及びシステム工学群), 応用科学群, 准教授 (00545981)
八谷 如美 地方独立行政法人東京都立産業技術研究センター, 開発本部開発第二部バイオ応用技術グループ, 主任研究員 (30408075)
田岡 万悟 首都大学東京, 理工学研究科, 准教授 (60271160)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | プロテオミクス / アグリゲート / LC-MS / BOPｓ / トリプシン消化 / 質量分析 / 固相抽出 / レーザーカッター / ＴＲＩＭ３２ |
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| Outline of Final Research Achievements |
Protein aggregation causes severe damages to cell and tissue functions. In the present study, we developed a new proteomics sample preparation method that is applicable to minute amounts of aggregates including those isolated from cultured cell lines with laser microdissection systems. This method, which we call BOPs, combines two advantages of current techniques, (1) unbiased binding of reversed-phase polymeric microbeads to any type of protein and (2) enhanced trypsin digestion efficiency in CH3CN-aqueous solvent systems, into a single-tube workflow. BOPs can readily be adapted to a wide range of diluted or denaturant-contaminated samples with high sensitivity, reproducibility, and reliability. The availability of the present BOPs make it especially attractive for next-stage proteomics of rare and limited aggregates.
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Report
(4 results)
Research Products
(7 results)
