A mild procedure for separating polypeptide chains prior to immunoprecipitation and western blotting analysis

Abstract Conventional cleavage of linked polypeptide chains by heating in SDS can so alter molecular structure as to interfere with antibody binding, on which both immunoprecipitation and ‘western blotting’ depend. As an alternative, gentle treatment with acid at room temperature or at 0° C was effective in separating the α and β chains of human MHC Class II glycoprotein dimers and proved superior in terms of preservation of at least one labile epitope on the β chain.