title: Immunoprecipitation: Western blot for proteins of low abundance.
abstract: Combining the procedures of immunoprecipitation and immunoblotting can help overcome some of the limitations of each separate procedure. Immunoblotting can identify immunoprecipitated proteins more specifically and with higher sensitivity than nonspecific protein stains or autoradiography. Immunoprecipitation can enrich proteins of interest to improve sensitivity for detection when compared with immunoblotting of whole cell extracts. Recently, immunoprecipitation-blotting helped us characterize a new autoantibody, anti-p155, and to test for the presence of the autoantibody in patient sera to study its clinical associations. The procedure for immunoprecipitation-blotting, with specific reference to this autoantibody test ("reverse" immunoprecipitation-blotting), is reported here in detail.
authors.affiliation: Arthritis and Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA.
authors.forename: Edward P
authors.initials: EP
authors.lastname: Trieu
authors.forename: Ira N
authors.initials: IN
authors.lastname: Targoff
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D057230:Limit of Detection; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D012685:Sepharose / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000032:analysis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D011506:Proteins; D013205:Staphylococcal Protein A; D012685:Sepharose