"Golden blot"--detection of polyclonal and monoclonal antibodies bound to antigens on nitrocellulose by protein A-gold complexes.
Protein A bound to particles of colloidal gold is widely used for cytochemical localization of antigens. A method for detecting immune complexes on "Western blots" with protein A-gold has been developed. It involved the following steps: (i) electrophoretic separation of proteins, (ii) transfer to nitrocellulose, (iii) incubation with antibodies, and (iv) staining with protein A-gold. The amount of antigen detectable was 10-50 ng. The sensitivity could be improved by subsequent photochemical silver staining (1-10 ng detected), and was then comparable to autoradiograms using iodinated protein A.
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000941:Antigens / Q000032:analysis*; D003101:Collodion; D006046:Gold; D006801:Humans; D007120:Immunochemistry; D010777:Photochemistry; D011485:Protein Binding; D013194:Staining and Labeling; D013205:Staphylococcal Protein A; D013552:Swine
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000941:Antigens; D013205:Staphylococcal Protein A; D006046:Gold; D003101:Collodion