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Development of a novel inspection method to observe intracellular molecular movement : an application of fluorescence correlation spectroscopy(FCS)

Research Project

Project/Area Number 22790531
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Laboratory medicine
Research InstitutionTokyo Women's Medical University

Principal Investigator

TANAKA Shotaro  東京女子医科大学, 医学部, 助教 (90380667)

Project Period (FY) 2010 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2010: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords蛍光相関分光法(FCS) / マイクロインジェクション / モノクローナル抗体 / 抗体 / protein 4.1R / 臨床検査医学 / PKCβ
Research Abstract

Anti-GFP mouse monoclonal antibody was labeled specifically on Fc region by red fluorescent dye. The antibody was injected into HeLa cells that stably expressed PKCβ-GFP. The formation of specific interaction between the antibody and PKCβ-GFP in the cells was demonstrated by analyzing with fluorescence cross-correlation spectroscopy(FCCS), but the interaction completely inhibited the translocation function of PKCβto plasma membrane. To avoid the formation of antibody cross-linking, a Fab fragment was prepared from anti-protein 4.1R antibody expressed by hybridoma, as a new candidate of the present study. Now the antibody function of the Fab fragment is being verified in the cells.

Report

(3 results)
  • 2011 Annual Research Report   Final Research Report ( PDF )
  • 2010 Annual Research Report
  • Research Products

    (6 results)

All 2012 2011

All Journal Article (2 results) (of which Peer Reviewed: 2 results) Presentation (4 results)

  • [Journal Article] Intracellular interactions between protein 4.1 and glycophorin C on transport vesicles, as determined by fluorescence correlation spectroscopy2012

    • Author(s)
      Tanaka S, Takakuwa Y, 計2名
    • Journal Title

      FEBS letters

      Volume: 586 Pages: 668-674

    • Related Report
      2011 Final Research Report
    • Peer Reviewed
  • [Journal Article] Intracellular interactions between protein 4.1 and glycophorin C on transport vesicles, as determined by fluorescence correlation spectroscopy Reference2012

    • Author(s)
      Shotaro Tanaka, Yuichi Takakuwa
    • Journal Title

      FEBS LETTERS

      Volume: 586 Pages: 668-674

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Presentation] 新しい細胞内分子診断法の開発-蛍光相関分光法(FCS)の臨床検査への応用-2011

    • Author(s)
      田中正太郎、高桑雄一
    • Organizer
      第84回日本生化学会大会
    • Place of Presentation
      京都
    • Year and Date
      2011-09-23
    • Related Report
      2011 Final Research Report
  • [Presentation] 新しい細胞内分子診断法の開発~蛍光相関分光法(FCS)の臨床検査への応用~2011

    • Author(s)
      田中正太郎
    • Organizer
      日本生化学会
    • Place of Presentation
      京都・京都国際会議場
    • Year and Date
      2011-09-23
    • Related Report
      2011 Annual Research Report
  • [Presentation] Observation of interaction between membrane skeletal protein and transmembrane protein on transporting vesicle and plasma membranes using fluorescence correlation spectroscopy (FCS)2011

    • Author(s)
      Shotaro Tanaka
    • Organizer
      日本細胞生物学会
    • Place of Presentation
      北海道・北海道大学
    • Year and Date
      2011-06-28
    • Related Report
      2011 Annual Research Report
  • [Presentation] 膜骨格タンパク質protein 4.1は輸送小胞を介して細胞膜へ輸送される:蛍光相関分光法(FCS)を用いた観察2011

    • Author(s)
      田中正太郎
    • Organizer
      日本膜学会
    • Place of Presentation
      東京・産業技術総合研究所
    • Year and Date
      2011-05-13
    • Related Report
      2011 Annual Research Report

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Published: 2010-08-23   Modified: 2016-04-21  

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