Chapter title |
Blue Native PAGE: Applications to Study Peroxisome Biogenesis
|
---|---|
Chapter number | 18 |
Book title |
Peroxisomes
|
Published in |
Methods in molecular biology, April 2017
|
DOI | 10.1007/978-1-4939-6937-1_18 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6935-7, 978-1-4939-6937-1
|
Authors |
Kanji Okumoto, Shigehiko Tamura, Yukio Fujiki |
Editors |
Michael Schrader |
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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