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Peroxisomes pp 197–205Cite as

Blue Native PAGE: Applications to Study Peroxisome Biogenesis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1595))

Abstract

Blue native polyacrylamide gel electrophoresis (BN-PAGE) is one of the useful methods to isolate protein complexes including membrane proteins under native conditions. In BN-PAGE, Coomassie Brilliant Blue G-250 binds to proteins and provides a negative charge for the electrophoretic separation without denaturing at neutral pH, allowing the analysis of molecular mass, oligomeric state, and composition of native protein complexes. BN-PAGE is widely applied to the characterization of soluble protein complexes as well as isolation of membrane protein complexes from biological membranes such as the complexes I–V of the mitochondrial respiratory chain and subcomplexes of the mitochondrial protein import machinery. BN-PAGE has also been introduced in the field of peroxisome research, for example, analysis of translocation machinery for peroxisomal matrix proteins embedded in the peroxisomal membrane. Here, we describe a basic protocol of BN-PAGE and its application to the study of peroxisome biogenesis.

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Acknowledgments

This work was supported in part by Grants-in-Aid for Scientific Research (24247038, 25112518, 25116717, 26116007, and 15 K14511 to Y.F.; 26440032 to K.O.; 24570134 to S.T.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and grants from the Takeda Science Foundation, the Naito Foundation, and the Japan Foundation for Applied Enzymology.

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Authors and Affiliations

  1. Department of Biology, Faculty of Sciences, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan

    Kanji Okumoto

  2. Division for Experimental Natural Science, Faculty of Arts and Science, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan

    Shigehiko Tamura

  3. Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Yukio Fujiki

Authors
  1. Kanji Okumoto
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  2. Shigehiko Tamura
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  3. Yukio Fujiki
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Corresponding author

Correspondence to Yukio Fujiki .

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Editors and Affiliations

  1. College of Life and Environmental Sciences, Biosciences, University of Exeter, Exeter, United Kingdom

    Michael Schrader

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Okumoto, K., Tamura, S., Fujiki, Y. (2017). Blue Native PAGE: Applications to Study Peroxisome Biogenesis. In: Schrader, M. (eds) Peroxisomes. Methods in Molecular Biology, vol 1595. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6937-1_18

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  • DOI: https://doi.org/10.1007/978-1-4939-6937-1_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6935-7

  • Online ISBN: 978-1-4939-6937-1

  • eBook Packages: Springer Protocols

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