Gas-phase Microsequencing of Peptides and Proteins with a Fluorescent Edman-type Reagent, Fluorescein Isothiocyanate
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概要
- 論文の詳細を見る
A fluorescent Edman-type reagent, fluorescein isothiocyanate (FITC), was adapted to a gas-phase sequencer fitted with a miniature reaction chamber. Fluorescein thiohydantoin amino acids were identified by on-line gradient HPLC with fluorescence monitoring. The optimized protocol for the coupling reaction using FITC and phenylisothiocyanate, and the degree of washing to remove the excess reagents were established. Myoglobin (5 pmol) and lysozyme (5 pmol) were analyzed by the sequencer to obtain a 35-51% initial yield (I. Y. ) and 82-88% repetitive yield (R. Y. ) for the former, and 51-66% I. Y. and 91-92% R. Y. for the latter. These figures permitted 20-25 amino acid residues to be identified from the N-terminus of 5 pmol samples applied to the sequencer.
- 社団法人日本農芸化学会の論文
- 1994-02-23
著者
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KAMIYA Hisao
Department of Marine Biochemistry, School of Marine Sciences, Kitasato University
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村本 光二
Lab. Of Functional Biomolecules Graduate School Of Life Sciences Tohoku Univ.
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Kamiya H
Department Of Marine Biochemistry School Of Marine Sciences Kitasato University
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Muramoto Koji
Laboratory Of Applied Phytochemistry Tohoku University Graduate School Of Agricultural Science
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Muramoto Koji
School Of Fisheries Sciences Kitasato University
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NOKIHARA Kiyoshi
Biotechnology Instruments Department, Shimazu Corporation
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Ueda A
Hiroshima Women's Univ. Hiroshima Jpn
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Kamiya H
Department Of Marine Biochemistry School Of Fisheries Sciences Kitasato University
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Ueda Akira
School of Fisheries Sciences, Kitasato University
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Kamiya Hisao
Tokyo University of Agriculture and Technology
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Nokihara K
Hipep Laboratories
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Nokihara Kiyoshi
Biotechnology Instruments Department Shimazu Corporation
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