Western Blotting: Remembrance of Things Past
Western blotting sprung from the need to develop a sensitive visual assay for the antigen specificity of monoclonal antibodies. The technique employed SDS-PAGE of protein antigens, electrophoretic replica transfer of gel-resolved proteins to unmodified nitrocellulose sheets, probing the immobilized antigens with hybridomas, and detection of antibody–antigen complexes with radiolabeled staphylococcal protein A and autoradiography. The simplicity and relevance of the method has led to its expansive application as an immunodiagnostic and a ubiquitous research tool in biology and medicine.
Key wordsWestern blotting SDS-PAGE Electroblotting Unmodified nitrocellulose Immobilized replica Antigen specificity Antibody–antigen complex Protein A
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