Detection of toxoplasma membrane antigens transferred from SDS-polyacrylamide gel to nitrocellulose with monoclonal antibody and avidin-biotin, peroxidase anti-peroxidase and immunoperoxidase methods.
Toxoplasma (Tp) membrane antigens separated by discontinuous SDS-polyacrylamide gel electrophoresis were electrophoretically transferred to nitrocellulose and detected with avidin-biotin (AB), peroxidase anti-peroxidase complex (PAP) or indirect immunoperoxidase (IIP) methods. In the AB method, the nitrocellulose was treated with biotinylated monoclonal antibodies and avidin-labeled peroxidase. In the PAP method, it was treated with monoclonal antibody, rabbit anti-mouse IgG antibody, goat anti-rabbit IgG antibody and PAP. Of the two, the AB method was the more sensitive and specific for Tp membrane antigen. The PAP method was less sensitive, but did not require chemical manipulation of the antibodies and was convenient and useful for analyzing Tp membrane antigens. The IIP method was more convenient, but had a lower sensitivity than the other methods.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology; D001360:Avidin; D001710:Biotin; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007124:Immunoenzyme Techniques / Q000592:standards; D051379:Mice; D008970:Molecular Weight; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate; D014122:Toxoplasma / Q000276:immunology; D014124:Toxoplasmosis, Animal / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D001360:Avidin; D012967:Sodium Dodecyl Sulfate; D001710:Biotin; D003101:Collodion