Solubilization of plasma membranes in anionic, non-ionic and zwitterionic surfactants for iso-dalt analysis: a critical evaluation

J Chromatogr. 1984 Sep 7;299(1):57-72. doi: 10.1016/s0021-9673(01)97821-5.

Abstract

A critical study has been made of the solubilizing properties of anionic, nonionic and zwitterionic surfactants to be used in the first dimension of two-dimensional isoelectric focusing-sodium dodecyl sulphate (IF-SDS) maps. Excess of SDS is a powerful solubilizing agent, but prevents proteins from entering the IF gel. Nonidet P-40 (NP-40)-urea mixtures are mediocre solubilizing agents, but are compatible with the IF dimension. Zwitterionic detergents (zwittergents) appear to exhibit a well balanced solubilizing power and are able to generate sharp two-dimensional maps, with round spots and minimal vertical and horizontal streaking. SB3-12 and SB3-14 appear to have the best solubilizing properties; shorter surfactants (SB3-8) exhibit a poor solubilization efficiency, while longer detergents (SB3-16) bind too strongly to hydrophobic regions in proteins. The random performance of non-ionic detergents has been attributed to their ability to form mixed micelles with the carrier ampholytes used in the IF step: depending on the relative ratio of NP-40 to Ampholines, different types of micelles would be formed, which, when reaching appropriate stoichiometries and charge densities, would mimic the behaviour of natural zwittergents. Acceptable two-dimensional maps can be obtained when the sample is lysed in limiting amounts of SDS (SDS:protein = 1:3), suggesting that excess of free SDS in solution is deleterious to the IF process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / analysis*
  • Detergents* / analysis
  • Electrophoresis, Polyacrylamide Gel / methods
  • Hydrogen-Ion Concentration
  • Isoelectric Focusing
  • L Cells
  • Mice
  • Quaternary Ammonium Compounds*
  • Sodium Dodecyl Sulfate
  • Surface-Active Agents* / analysis

Substances

  • Detergents
  • Quaternary Ammonium Compounds
  • Surface-Active Agents
  • zwittergent 3-12
  • Sodium Dodecyl Sulfate