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Native immunoblotting of blue native gels to identify conformation‐specific antibodies |
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| Authors: | Volker Zickermann Zibiernisha Wumaier Blanka Wrzesniewska Carola Hunte Hermann Schägger |
| Affiliation: | 1. Goethe‐Universit?t, Fachbereich Medizin, Molekulare Bioenergetik, Cluster of Excellence Frankfurt, “Macromolecular Complexes”, Center for Membrane Proteomics, Frankfurt am Main, Germany;2. Institute of Membrane and Systems Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK |
| Abstract: | A large repertoire of immunological methods permits monitoring the interaction of antibodies with their specific antigen. However, recognition of a protein by a conformation‐specific antibody represents a challenge because native conditions must be kept throughout the assay. Native immunoblotting of blue native gels conserves the native state by using Tween 20 instead of methanol for the obligatory destaining of the blot membrane. We validate the new technique with a set of monoclonal antibodies against respiratory NADH:ubiquinone oxidoreductase. |
| Keywords: | Blue native electrophoresis Monoclonal antibody Native immunoblot NADH dehydrogenase Technology Surface plasmon resonance |