Haptoglobin subtyping by isoelectric focusing in miniaturized polyacrylamide gels rehydrated in presence of 2-mercaptoethanol.
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.
Alonso
A
A
Instituto Nacional de Toxicologia, Madrid.
Visedo
G
G
Sancho
M
M
Fernández-Piqueras
J
J
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
D000180:Acrylic Resins; D006242:Haptoglobins; C016680:polyacrylamide gels; D008623:Mercaptoethanol; D014508:Urea; D009439:Neuraminidase