A mild procedure for separating polypeptide chains prior to immunoprecipitation and western blotting analysis.
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 degrees C was effective in separating the alpha and beta chains of human MHC Class II glycoprotein dimers and proved superior in terms of preservation of at least one labile epitope on the beta chain.
D000937:Antigen-Antibody Reactions; D002460:Cell Line; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D006680:HLA Antigens / Q000032:analysis*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010209:Paper; D010455:Peptides / Q000032:analysis* / Q000276:immunology; D011233:Precipitin Tests; D012967:Sodium Dodecyl Sulfate; D013696:Temperature; D013997:Time Factors
D006680:HLA Antigens; D010455:Peptides; D012967:Sodium Dodecyl Sulfate; D003101:Collodion