title: Haptoglobin subtyping by isoelectric focusing in miniaturized polyacrylamide gels rehydrated in presence of 2-mercaptoethanol.
abstract: A fast isoelectric focusing method for routine haptoglobin (Hp) subtyping is presented. This method is based on isoelectric focusing, under reducting conditions, of neuraminidase-treated plasma samples by using dry miniaturized (interelectrode distance: 55 mm) polyacrylamide gel, rehydrated in presence of 2-mercaptoethanol and a mixture of pharmalyte carrier ampholytes (pH 4-6.5 and pH 6-8) followed by immunoblotting. The presence of 2-mercaptoethanol in the gel prevented refolding of the Hp alpha and Hp beta chains during focusing, making it possible to obtain a sharp Hp band pattern with a clear separation of the different Hp alpha allelic products (1S, 1F, 2FS, 2SS and 2FF). A population study carried out with 250 unrelated individuals living in Central Spain is also presented.
authors.affiliation: Instituto Nacional de Toxicologia, Madrid.
authors.forename: A
authors.initials: A
authors.lastname: Alonso
authors.forename: G
authors.initials: G
authors.lastname: Visedo
authors.forename: M
authors.initials: M
authors.lastname: Sancho
authors.forename: J
authors.initials: J
authors.lastname: Fernández-Piqueras
mesh_terms: D000180:Acrylic Resins; D000483:Alleles; D005060:Europe; D005787:Gene Frequency; D006242:Haptoglobins / Q000145:classification* / Q000235:genetics; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D008623:Mercaptoethanol; D009439:Neuraminidase; D010641:Phenotype; D013030:Spain; D014508:Urea
chemical_list: D000180:Acrylic Resins; D006242:Haptoglobins; C016680:polyacrylamide gels; D008623:Mercaptoethanol; D014508:Urea; D009439:Neuraminidase