A simple method for peptide purification as a basis for peptidome analysis
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概要
- 論文の詳細を見る
We developed a simple method for purifying peptides from crude extracts of animal tissues. Peptides free of high-molecular-mass (HMM) proteins were prepared by differential solubilization of proteins and peptides followed by tricine-SDS-PAGE. Proteins and peptides in a tissue extract were co-precipitated by slowly dropping the extracts into acetone at −40°C. The precipitates were dissolved in 2 mM DTT and separated by centrifugation. The peptide-enriched supernatant was resolved by tricine-SDS-PAGE and portions of the lanes below 10 kDa were excised. Peptides were extracted from the gel pieces in 40% acetonitrile and fractionated by two-step reversed-phase HPLC. We evaluated the method by extracting peptides from the livers and kidneys of rats and diabetic model mice. The HPLC elution profiles of peptide mixtures were reproducible, peptide recovery from tricine-SDS-PAGE gels was high and we identified one HPLC-fractionated peptide by amino-acid sequencing. The present method should serve as a good foundation for future peptidomics.
- 2005-03-01
著者
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Fukutomi Toshiyuki
Department of Pharmacology and Toxicology, Kyorin University School of Medicine, Japan
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MAEDA Tadakazu
Department of Physics, School of Science
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Maeda Tadakazu
Laboratory Of Biomolecular Dynamics Department Of Physics Kitasato University School Of Science
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Maeda Tadakazu
Graduate School Of Science Kitasato University
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Maeda Tadakazu
Department Of Physics Faculty Of Science Nagoya University:mitsubishi-kasei Institute Of Life Scienc
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KODERA Yoshio
Departments of Physics, Kitasato University School of Science
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Fukutomi Toshiyuki
Laboratory Of Biomolecular Dynamics Department Of Physics Kitasato University School Of Science
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Fukutomi Toshiyuki
Department Of Pharmacology And Toxicology Kyorin University School Of Medicine
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Omori Akira
Mitsubishi Kagaku Institute Of Life Science (mitils)
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Kodera Yoshio
Laboratory Of Biomolecular Dynamics Department Of Physics Kitasato University School Of Science
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FURUDATE Sen-ichi
Department of Laboratory Animal Science, Kitasato University School of Medicine
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Matsui Takashi
Department Of Physics School Of Science Kitasato University
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Furudate S
Department Of Laboratory Animal Science School Of Medicine Kitasato University
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Furudate Sen-ichi
Department Of Laboratory Animal Science Kitasato University School Of Medicine
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Kodera Yoshio
Department Of Physics Kitasato University School Of Science
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KOGO Tomihisa
Department of Physics, Kitasato University School of Science
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Maeda Tadakazu
Department Of Physics Kitasato University School Of Science
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Kogo Tomihisa
Department Of Physics Kitasato University School Of Science
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