| Project/Area Number |
20310121
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied genomics
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| Research Institution | Yokohama City University |
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Principal Investigator |
HIRANO Hisashi Yokohama City University, 生命ナノシステム科学研究科, 教授 (00275075)
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| Co-Investigator(Kenkyū-buntansha) |
KAWASAKI HIROSHI 横浜市立大学, 生命ナノシステム科学研究科, 准教授 (70169704)
ARAKAWA NORIAKI 横浜市立大学, 生命ナノシステム科学研究科, 助教 (60398394)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥19,500,000 (Direct Cost: ¥15,000,000、Indirect Cost: ¥4,500,000)
Fiscal Year 2010: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2009: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2008: ¥10,790,000 (Direct Cost: ¥8,300,000、Indirect Cost: ¥2,490,000)
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| Keywords | タンパク質 / 翻訳後修飾 / プロテインチップ / 質量分析装置 / 癌 |
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| Research Abstract |
In the 1990s, techniques were developed to transfer proteins from gels onto PVDF membranes, and to identify the proteins on the membranes by MALDI-MS. However, the low ionization efficiency of peptides and proteins immobilized on the membranes often renders these techniques useless. Therefore, it is essential to develop membranes on which peptides and proteins can be efficiently ionized for MS analysis. In this study, we found that 0.1-μm pores PVDF membranes can be used for highly efficient and effective electroblotting and MALDI-MS analysis. This membrane showed high ion yield in MALDI-MS analysis of peptides compared to conventional PVDF membranes. In all, 38 yeast proteins with various physicochemical characteristics were separated by 2-DE, immobilized onto the 0.1-μm pore PVDF membranes, and successfully identified by MALDI-MS/MS.
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