High-resolution Native-PAGE for membrane proteins capable of fluorescence detection and hydrodynamic state evaluation

Makoto Ihara, Noriko Matsuura, Atsuko Yamashita

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

An improved native polyacrylamide gel electrophoresis (PAGE) method capable of evaluating the hydrodynamic states of membrane proteins and allowing in-gel fluorescence detection was established. In this method, bis(alkyl) sulfosuccinate is used to provide negative charges for detergent-solubilized membrane proteins to facilitate proper electrophoretic migration without disturbing their native hydrodynamic states. The method achieved high-resolution electrophoretic separation, in good agreement with the elution profiles obtained by size exclusion chromatography. The applicability of in-gel fluorescence detection for tagged green fluorescent protein (GFP) facilitates the analysis of samples without any purification. This method might serve as a general analytical technique for assessing the folding, oligomerization, and protein complex formation of membrane proteins.

Original languageEnglish
Pages (from-to)217-223
Number of pages7
JournalAnalytical Biochemistry
Volume412
Issue number2
DOIs
Publication statusPublished - May 15 2011
Externally publishedYes

Fingerprint

Native Polyacrylamide Gel Electrophoresis
Hydrodynamics
Electrophoresis
Membrane Proteins
Fluorescence
Gels
Oligomerization
Size exclusion chromatography
Green Fluorescent Proteins
Detergents
Purification
Protein Folding
Gel Chromatography
polyacrylamide gels
Proteins

Keywords

  • Fluorescence detection size exclusion chromatography (FSEC)
  • Green fluorescent protein (GFP)
  • Hydrodynamic states
  • Membrane proteins
  • Native PAGE

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

Cite this

High-resolution Native-PAGE for membrane proteins capable of fluorescence detection and hydrodynamic state evaluation. / Ihara, Makoto; Matsuura, Noriko; Yamashita, Atsuko.

In: Analytical Biochemistry, Vol. 412, No. 2, 15.05.2011, p. 217-223.

Research output: Contribution to journalArticle

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