General detection of proteins after electroblotting by trinitrobenzene sulphonic acid derivatization and immunochemical staining with a monoclonal antibody against the trinitrophenyl group.
A sensitive method for the general detection of proteins electroblotted onto nitrocellulose sheets after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. The proteins on the blots were reacted with 2,4,6-trinitrobenzene sulphonic acid. The resulting trinitrophenyl groups on the proteins were rendered visible by immunochemical staining with a monoclonal anti-trinitrophenyl antibody, and a peroxidase-conjugated second antibody. Using various proteins, the method was compared to the amidoblack method for staining of protein blots. The method was 10-100-fold more sensitive than the amidoblack method. Amounts as low as 1 ng of human serum albumin could be detected.
D000911:Antibodies, Monoclonal / Q000276:immunology; D004586:Electrophoresis / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D014301:Trinitrobenzenes / Q000276:immunology; D014302:Trinitrobenzenesulfonic Acid
D000911:Antibodies, Monoclonal; D011506:Proteins; D014301:Trinitrobenzenes; D014302:Trinitrobenzenesulfonic Acid