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Native immunoblotting of blue native gels to identify conformation‐specific antibodies
Author(s) -
Zickermann Volker,
Wumaier Zibiernisha,
Wrzesniewska Blanka,
Hunte Carola,
Schägger Hermann
Publication year - 2009
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200900468
Subject(s) - native state , antibody , western blot , monoclonal antibody , chemistry , antigen , biochemistry , microbiology and biotechnology , biology , immunology , gene
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.