Non-gradient blue native polyacrylamide gel electrophoresis

Xiaoting Luo, Jinzi Wu, Zhen Jin, Liang Jun Yan

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Gradient blue native polyacrylamide gel electrophoresis (BN-PAGE) is a well established and widely used technique for activity analysis of high-molecularweight proteins, protein complexes, and protein-protein interactions. Since its inception in the early 1990s, a variety of minor modifications have been made to this gradient gel analytical method. Here we provide a major modification of the method, which we call non-gradient BN-PAGE. The procedure, similar to that of non-gradient SDS-PAGE, is simple because there is no expensive gradient maker involved. The non-gradient BN-PAGE protocols presented herein provide guidelines on the analysis of mitochondrial protein complexes, in particular, dihydrolipoamide dehydrogenase (DLDH) and those in the electron transport chain. Protocols for the analysis of blood esterases or mitochondrial esterases are also presented. The non-gradient BN-PAGE method may be tailored for analysis of specific proteins according to their molecular weight regardless of whether the target proteins are hydrophobic or hydrophilic.

Original languageEnglish
Pages (from-to)19.29.1-19.29.12
JournalCurrent Protocols in Protein Science
Volume2017
DOIs
StatePublished - 1 Feb 2017

Fingerprint

Native Polyacrylamide Gel Electrophoresis
Electrophoresis
Proteins
Esterases
Dihydrolipoamide Dehydrogenase
Mitochondrial Proteins
Electron Transport
polyacrylamide gels
Polyacrylamide Gel Electrophoresis
Blood
Molecular Weight
Gels
Molecular weight
Guidelines

Keywords

  • Blue native gel
  • Electrophoresis
  • Esterase
  • Mitochondria
  • Nongradient

Cite this

Luo, Xiaoting ; Wu, Jinzi ; Jin, Zhen ; Yan, Liang Jun. / Non-gradient blue native polyacrylamide gel electrophoresis. In: Current Protocols in Protein Science. 2017 ; Vol. 2017. pp. 19.29.1-19.29.12.
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Non-gradient blue native polyacrylamide gel electrophoresis. / Luo, Xiaoting; Wu, Jinzi; Jin, Zhen; Yan, Liang Jun.

In: Current Protocols in Protein Science, Vol. 2017, 01.02.2017, p. 19.29.1-19.29.12.

Research output: Contribution to journalArticle

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AU - Jin, Zhen

AU - Yan, Liang Jun

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