title: Isolation of smooth-type lipopolysaccharides to electrophoretic homogeneity.
abstract: Polyacrylamide slab gel electrophoresis in the presence of sodium dodecyl sulfate or sodium deoxycholate (SDS- or DOC-slab-PAGE) is a powerful technique for the separation of smooth(S)-type bacterial lipopolysaccharides (LPS). In order to recover the individual LPS species from the polyacrylamide gel for subsequent analyses, a sensitive, nondestructive reverse staining of slab-PAGE-separated LPS has been developed. The individual reverse-stained LPS bands can be rapidly and efficiently recovered into an aqueous 5% triethylamine solution when they are extruded to produce fine gel microparticles. Based on these principles, an isolation methodology that combines preparative slab-PAGE, reverse staining, extrusion, and passive elution can be used to isolate, to electrophoretic homogeneity, micrograms to hundreds of micrograms of individual LPS species successfully from smooth-type LPS mixtures.
authors.affiliation: Department of Vaccinology, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands. elder.pupo@yahoo.com
authors.forename: Elder
authors.initials: E
authors.lastname: Pupo
mesh_terms: D003840:Deoxycholic Acid; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005021:Ethylamines / Q000378:metabolism; D008070:Lipopolysaccharides / Q000032:analysis* / Q000302:isolation & purification*; D016622:Silver Staining / Q000379:methods; D012967:Sodium Dodecyl Sulfate
chemical_list: D005021:Ethylamines; D008070:Lipopolysaccharides; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; C016162:triethylamine