title: High resolution isoelectric focusing of immunoprecipitated proteins under denaturing conditions. A simple analytical method applied to the study of complement component polymorphisms.
abstract: A simple analytical method for the study of structural protein polymorphisms is described. It consists of the immunoprecipitation of non-radiolabeled proteins using monospecific polyclonal antibodies followed by isoelectric focusing (IEF) under completely denaturing conditions in vertical polyacrylamide slab gels. The method uses small amounts of sample (usually unfractionated plasma or serum), requires no sophisticated equipment and allows the screening of large numbers of samples with comparatively small effort. This method has been applied in the identification of 2 human complement-component polymorphisms, C4-binding protein (C4-bp) and factor H (beta 1H).
mesh_terms: D000937:Antigen-Antibody Reactions; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003180:Complement C3b Inactivator Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D017242:Complement Factor H; D003165:Complement System Proteins / Q000235:genetics*; D005796:Genes; D006801:Humans; D016167:Integrin alphaXbeta2; D007525:Isoelectric Focusing / Q000379:methods*; D008970:Molecular Weight; D011110:Polymorphism, Genetic; D011234:Precipitins / Q000032:analysis*; D011489:Protein Denaturation
chemical_list: C493495:CFH protein, human; D002352:Carrier Proteins; D003180:Complement C3b Inactivator Proteins; D016167:Integrin alphaXbeta2; D011234:Precipitins; D017242:Complement Factor H; D003165:Complement System Proteins