Two dimensional electrophoresis and immunoblotting for the study of ovine whey protein polymorphism.

Abstract

Different electrophoretic techniques have been applied to the study of ovine whey protein polymorphism. Isoelectric focusing (IEF) over the pH gradient 3.5-9.5 was a suitable method for resolving both genetic variants of ovine beta-lactoglobulin (beta-lg); using this type of IEF, ovine beta-lg A seemed to be defined by a major and a minor 'satellite' band. Approximate isoelectric point, relative molecular mass, amino acid composition and N-terminal sequence of this minor band were determined. A good separation of ovine beta-lg was achieved using ultrathin layer isoelectric focusing (UTLIEF) over the pH gradient 2.5-7.0. However, addition of urea in UTLIEF gels led to some differences in the patterns of the ovine whey proteins when compared with those obtained by IEF on gels not containing urea. Two dimensional electrophoretic techniques provided a considerable separation of ovine whey proteins. Immunoblotting applied to a two dimensional separation permitted the identification of the bands belonging to beta-lg and alpha-lactalbumin fractions.