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Western Blotting: Remembrance of Things Past

  • W. Neal Burnette
Part of the Methods in Molecular Biology book series (MIMB, volume 536)

Summary

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 words

Western blotting SDS-PAGE Electroblotting Unmodified nitrocellulose Immobilized replica Antigen specificity Antibody–antigen complex Protein A 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Molecular Pharmaceutics CorporationChapel HillUSA

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