Two-dimensional electrophoresis of membrane proteins using immobilized pH gradients

Mark P. Molloy*

*Corresponding author for this work

Research output: Contribution to journalReview article

273 Citations (Scopus)

Abstract

Two-dimensional electrophoresis (2-DE) is a highly resolving technique for arraying proteins by isoelectric point and molecular mass. To date, the resolving ability of 2-DE for protein separation is unsurpassed, thus ensuring its use as the fundamental separation method for proteomics. When immobilized pH gradients (IPGs) are used for isoelectric focusing in the first dimension, excellent reproducibility and high protein load capacity can be achieved. While this has been beneficial for separations of soluble and mildly hydrophobic proteins, separations of membrane proteins and other hydrophobic proteins with IPGs have often been poor. Stimulated by the growing interest in proteomics, recent developments in 2-DE methodology have been aimed at rectifying this situation. Improvements have been made in the area of protein solubilization and sample fractionation, leading to a revamp of traditional approaches for 2-DE of membrane proteins. This review explores these developments. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)1-10
Number of pages10
JournalAnalytical Biochemistry
Volume280
Issue number1
DOIs
Publication statusPublished - 10 Apr 2000

Keywords

  • Extraction
  • Immobilized pH gradients
  • Membrane proteins
  • Proteomics
  • Solubilization
  • Two-dimensional electrophoresis

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