Haptoglobin subtyping by isoelectric focusing in miniaturized polyacrylamide gels rehydrated in presence of 2‐mercaptoethanol

Publisher Website
Google Scholar
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, rehydráted 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 α and Hp β chains during focusing, making it possible to obtain a sharp Hp band pattern with a clear separation of the different Hp α allelic products (1S, 1F, 2FS, 2SS and 2FF). A population study carried out with 250 unrelated individuals living in Central Spain is also presented.