Generation and characterization of monoclonal antibodies that recognize human and murine supervillin protein isoforms
UMass Chan Affiliations
Department of Radiology, Division of Cell Biology and ImagingDocument Type
Journal ArticlePublication Date
2018-10-17Keywords
ImmunoprecipitationCloning
Immunohistochemistry techniques
Hybridomas
Monoclonal antibodies
Enzyme-linked immunoassays
Immunoblotting
Cell staining
Amino Acids, Peptides, and Proteins
Cancer Biology
Cell Biology
Enzymes and Coenzymes
Immunology and Infectious Disease
Neoplasms
Metadata
Show full item recordAbstract
Supervillin isoforms have been implicated in cell proliferation, actin filament-based motile processes, vesicle trafficking, and signal transduction. However, an understanding of the roles of these proteins in cancer metastasis and physiological processes has been limited by the difficulty of obtaining specific antibodies against these highly conserved membrane-associated proteins. To facilitate research into the biological functions of supervillin, monoclonal antibodies were generated against the bacterially expressed human supervillin N-terminus. Two chimeric monoclonal antibodies with rabbit Fc domains (clones 1E2/CPTC-SVIL-1; 4A8/CPTC-SVIL-2) and two mouse monoclonal antibodies (clones 5A8/CPTC-SVIL-3; 5G3/CPTC-SVIL-4) were characterized with respect to their binding sites, affinities, and for efficacy in immunoblotting, immunoprecipitation, immunofluorescence microscopy and immunohistochemical staining. Two antibodies (1E2, 5G3) recognize a sequence found only in primate supervillins, whereas the other two antibodies (4A8, 5A8) are specific for a more broadly conserved conformational epitope(s). All antibodies function in immunoblotting, immunoprecipitation and in immunofluorescence microscopy under the fixation conditions identified here. We also show that the 5A8 antibody works on immunohistological sections. These antibodies should provide useful tools for the study of mammalian supervillins.Source
PLoS One. 2018 Oct 17;13(10):e0205910. doi: 10.1371/journal.pone.0205910. eCollection 2018. Link to article on publisher's site
DOI
10.1371/journal.pone.0205910Permanent Link to this Item
http://hdl.handle.net/20.500.14038/40821PubMed ID
30332471Related Resources
Rights
Copyright: © 2018 Smith et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Distribution License
http://creativecommons.org/licenses/by/4.0/ae974a485f413a2113503eed53cd6c53
10.1371/journal.pone.0205910
Scopus Count
Except where otherwise noted, this item's license is described as Copyright: © 2018 Smith et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.