Electroimmunoassay-immunoblotting (EIA-IB) for the utilization of monoclonal antibodies in quantitative immunoelectrophoresis: the method and its applications.
Immunoprecipitates formed in conventional electroimmunoassays were solubilized by brief immersion of the agarose gels in sodium dodecylsulfate. The proteins were then transferred to nitrocellulose sheets by electroblotting. The blotted proteins were readily amenable to analyses by non-precipitating monoclonal antibodies and the immunoblots were developed with second antibody/biotin-streptavidin-peroxidase staining. The electroimmunoassay-immunoblot (EIA-IB) method is of value in (1) specificity assays of monoclonal antibodies in crossed immunoelectrophoresis; (2) analysis of specific molecular interactions between proteins; (3) rapid screening and simple identification of monoclonal antibodies by line immunoelectrophoresis-immunoblotting.
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D003101:Collodion; D003182:Complement C5 / Q000276:immunology / Q000378:metabolism; D050776:Complement C5b; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007122:Immunoelectrophoresis / Q000379:methods*; D007123:Immunoelectrophoresis, Two-Dimensional / Q000379:methods*; D010209:Paper; D011817:Rabbits
D000911:Antibodies, Monoclonal; D003182:Complement C5; D050776:Complement C5b; D003101:Collodion