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 journalArticlepeer-review

7 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

Keywords

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

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'High-resolution Native-PAGE for membrane proteins capable of fluorescence detection and hydrodynamic state evaluation'. Together they form a unique fingerprint.

Cite this