『徳島大学 教育・研究者情報データベース (EDB)』---[学外] /
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種別 (必須): 学術論文 (審査論文) [継承]
言語 (必須): 英語 [継承]
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著者 (必須): 1.中村 教泰
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2. (英) Tsumoto Kouhei (日) (読)
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3.石村 和敬
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4. (英) Kumagai Izumi (日) (読)
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題名 (必須): (英) Phage library panning against cytosolic fraction of cells using quantitative dot blotting assay: application of selected VH to histochemistry.  (日)    [継承]
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要約 (任意): (英) Comprehensive preparations of antibodies against various kinds of proteins in cells would be useful in proteome research and antibody-based research. Here we report the panning of a human antibody heavy chain variable domain (VH) phage library against a cytosolic fraction of rat liver to obtain antibodies specific for certain cytoplasmic proteins. Rat liver specimens were homogenized and subjected to differential centrifugation. A 125000 x g supernatant (rat liver cytosol, RLC) was immobilized onto a nitrocellulose membrane and subjected to phage VH library panning. For efficient assessment of binding phages, we established a system that was a combination of monoclonal phage ELISA and quantitative dot blotting of phages. The VH genes of the binding phages were selected and expressed as VH--bacterial alkaline phosphatase (PhoA) conjugates (VH/RLC--PhoAs) in Escherichia coli. One of the VH/RLC--PhoAs stained one major band on Western blotting of RLC and also stained the cytoplasm of hepatocytes histochemically. This is the first report of phage library panning against the cytosolic fraction of cells to obtain human VH fragments, and the application of those human VH fragments to histochemical study.  (日)    [継承]
キーワード (推奨): 1. (英) Amino Acid Sequence (日) (読) [継承]
2. (英) Animals (日) (読) [継承]
3. (英) Base Sequence (日) (読) [継承]
4. (英) Blotting, Western (日) (読) [継承]
5. (英) Cytosol (日) (読) [継承]
6.DNA (DNA) [継承]
7. (英) Enzyme-Linked Immunosorbent Assay (日) (読) [継承]
8. (英) Humans (日) (読) [継承]
9. (英) Immunoblotting (日) (読) [継承]
10. (英) Immunoglobulin Heavy Chains (日) (読) [継承]
11. (英) Immunoglobulin Variable Region (日) (読) [継承]
12. (英) Liver (日) (読) [継承]
13. (英) Male (日) (読) [継承]
14. (英) Peptide Library (日) (読) [継承]
15. (英) Rats (日) (読) [継承]
16. (英) Rats, Wistar (日) (読) [継承]
発行所 (推奨):
誌名 (必須): Journal of Immunological Methods (Association of Medical Laboratory Immunologists)
(pISSN: 0022-1759, eISSN: 1872-7905)

ISSN (任意): 0022-1759
ISSN: 0022-1759 (pISSN: 0022-1759, eISSN: 1872-7905)
Title: Journal of immunological methods
Title(ISO): J Immunol Methods
Publisher: Elsevier BV
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年月日 (必須): 西暦 2002年 3月 1日 (平成 14年 3月 1日) [継承]
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DOI (任意): 10.1016/S0022-1759(01)00547-6    (→Scopusで検索) [継承]
PMID (任意): 11861066    (→Scopusで検索) [継承]
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Scopus (任意): 2-s2.0-0036498673 [継承]
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備考 (任意): 1.(英) Article.Affiliation: Department of Anatomy and Cell Biology, School of Medicine, Tokushima University, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.  (日)    [継承]
2.(英) Article.PublicationTypeList.PublicationType: Journal Article  (日)    [継承]
3.(英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't  (日)    [継承]

標準的な表示

和文冊子 ● Michihiro Nakamura, Kouhei Tsumoto, Kazunori Ishimura and Izumi Kumagai : Phage library panning against cytosolic fraction of cells using quantitative dot blotting assay: application of selected VH to histochemistry., Journal of Immunological Methods, Vol.261, No.1-2, 65-72, 2002.
欧文冊子 ● Michihiro Nakamura, Kouhei Tsumoto, Kazunori Ishimura and Izumi Kumagai : Phage library panning against cytosolic fraction of cells using quantitative dot blotting assay: application of selected VH to histochemistry., Journal of Immunological Methods, Vol.261, No.1-2, 65-72, 2002.

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