Fluorescence Labeling of a Cytokine with Desthiobiotin-Tagged Fluorescent Puromycin(METHODS)
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概要
- 論文の詳細を見る
Fluorescence labeling of a cytokine at a specific site is required for observing cytokine-receptor interactions in living cells at the single-molecule level. Here, we demonstrated the C-terminus-specific fluorescence labeling of histidine-tagged thrombopoietin(TPO), a ligand for Mpl, with desthiobiotin-tagged fluorescent puromycin. Fluorescent TPO, purified by tandem affinity purification, stimulated the proliferation of Mpl-expressing cells. Within 10 min of its addition, fluorescent TPO was found to be diffusely distributed on the cell membranes of Mpl-expressing cells, and gradually accumulated to form fluorescent spots. This method is applicable for studying the spatial and temporal distributions of cytokines in individual living cells.
- 2008-03-25
著者
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KATO Takashi
Graduate School of Science and Engineering, Waseda University
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Funatsu Takashi
Graduate School of Pharmaceutical Sciences, The University of Tokyo
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Yamagishi Mai
Graduate School Of Pharmaceutical Sciences The University Of Tokyo:cnbi
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Yamagishi Mai
Graduate School Of Pharmaceutical Sciences The University Of Tokyo
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Funatsu Takashi
Graduate School Of Pharmaceutical Sciences The University Of Tokyo
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Funatsu Takashi
Graduate School Of Pharmaceutical Sciences The University Of Tokyo:center For Nanobio Integration Th
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Sakamoto Akihiko
Graduate School of Pharmaceutical Sciences, The University of Tokyo
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Watanabe Takafumi
Graduate School of Advanced Science and Engineering, Waseda University
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Aizawa Youichi
Graduate School of Advanced Science and Engineering, Waseda University
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Kuribara Takeshi
Major In Integrative Bioscience And Biomedical Engineering Graduate School Of Science And Engineerin
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Watanabe Takafumi
Graduate School Of Advanced Science And Engineering Waseda University
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Aizawa Youichi
Graduate School Of Advanced Science And Engineering Waseda University
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Sakamoto Akihiko
Grad. Sch. Of Pharm. Sci. The Univ. Of Tokyo
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Kato Takashi
Fac. Of Ed. And Int. Arts And Sci. Waseda Univ.
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Kato Takashi
Graduate School For International Development And Cooperation Hiroshima University
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