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dc.contributor.authorSmith, Tara C.
dc.contributor.authorSaul, Richard G.
dc.contributor.authorBarton, Elisabeth R.
dc.contributor.authorLuna, Elizabeth J.
dc.date2022-08-11T08:09:51.000
dc.date.accessioned2022-08-23T16:46:08Z
dc.date.available2022-08-23T16:46:08Z
dc.date.issued2018-10-17
dc.date.submitted2018-11-29
dc.identifier.citation<p>PLoS One. 2018 Oct 17;13(10):e0205910. doi: 10.1371/journal.pone.0205910. eCollection 2018. <a href="https://doi.org/10.1371/journal.pone.0205910">Link to article on publisher's site</a></p>
dc.identifier.issn1932-6203 (Linking)
dc.identifier.doi10.1371/journal.pone.0205910
dc.identifier.pmid30332471
dc.identifier.urihttp://hdl.handle.net/20.500.14038/40821
dc.description.abstractSupervillin 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.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=30332471&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rightsCopyright: © 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.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectImmunoprecipitation
dc.subjectCloning
dc.subjectImmunohistochemistry techniques
dc.subjectHybridomas
dc.subjectMonoclonal antibodies
dc.subjectEnzyme-linked immunoassays
dc.subjectImmunoblotting
dc.subjectCell staining
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectCancer Biology
dc.subjectCell Biology
dc.subjectEnzymes and Coenzymes
dc.subjectImmunology and Infectious Disease
dc.subjectNeoplasms
dc.titleGeneration and characterization of monoclonal antibodies that recognize human and murine supervillin protein isoforms
dc.typeJournal Article
dc.source.journaltitlePloS one
dc.source.volume13
dc.source.issue10
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=4640&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/3628
dc.identifier.contextkey13391902
refterms.dateFOA2022-08-23T16:46:09Z
html.description.abstract<p>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.</p>
dc.identifier.submissionpathoapubs/3628
dc.contributor.departmentDepartment of Radiology, Division of Cell Biology and Imaging
dc.source.pagese0205910


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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.
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.