Blue native PAGE: Applications to study peroxisome biogenesis

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages197-205
Number of pages9
DOIs
Publication statusPublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1595
ISSN (Print)1064-3745

Fingerprint

Native Polyacrylamide Gel Electrophoresis
Peroxisomes
Proteins
Membrane Proteins
Membranes
Mitochondrial Proteins
Electron Transport
Research

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Okumoto, K., Tamura, S., & Fujiki, Y. (2017). Blue native PAGE: Applications to study peroxisome biogenesis. In Methods in Molecular Biology (pp. 197-205). (Methods in Molecular Biology; Vol. 1595). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6937-1_18

Blue native PAGE : Applications to study peroxisome biogenesis. / Okumoto, Kanji; Tamura, Shigehiko; Fujiki, Yukio.

Methods in Molecular Biology. Humana Press Inc., 2017. p. 197-205 (Methods in Molecular Biology; Vol. 1595).

Research output: Chapter in Book/Report/Conference proceedingChapter

Okumoto, K, Tamura, S & Fujiki, Y 2017, Blue native PAGE: Applications to study peroxisome biogenesis. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1595, Humana Press Inc., pp. 197-205. https://doi.org/10.1007/978-1-4939-6937-1_18
Okumoto K, Tamura S, Fujiki Y. Blue native PAGE: Applications to study peroxisome biogenesis. In Methods in Molecular Biology. Humana Press Inc. 2017. p. 197-205. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-6937-1_18
Okumoto, Kanji ; Tamura, Shigehiko ; Fujiki, Yukio. / Blue native PAGE : Applications to study peroxisome biogenesis. Methods in Molecular Biology. Humana Press Inc., 2017. pp. 197-205 (Methods in Molecular Biology).
@inbook{e5c9c3b9fccb444bac29c9ec3fc97555,
title = "Blue native PAGE: Applications to study peroxisome biogenesis",
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.",
author = "Kanji Okumoto and Shigehiko Tamura and Yukio Fujiki",
year = "2017",
month = "1",
day = "1",
doi = "10.1007/978-1-4939-6937-1_18",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "197--205",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Blue native PAGE

T2 - Applications to study peroxisome biogenesis

AU - Okumoto, Kanji

AU - Tamura, Shigehiko

AU - Fujiki, Yukio

PY - 2017/1/1

Y1 - 2017/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85017578165&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85017578165&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-6937-1_18

DO - 10.1007/978-1-4939-6937-1_18

M3 - Chapter

T3 - Methods in Molecular Biology

SP - 197

EP - 205

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -