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dc.creatorNath, S
dc.creatorSpencer, VA
dc.creatorHan, J
dc.creatorChang, H
dc.creatorZhang, K
dc.creatorFontenay, GV
dc.creatorAnderson, C
dc.creatorHyman, JM
dc.creatorNilsen-Hamilton, M
dc.creatorChang, YT
dc.creatorParvin, B
dc.date.accessioned2021-01-31T22:39:43Z
dc.date.available2021-01-31T22:39:43Z
dc.date.issued2012-01-05
dc.identifier.issn1932-6203
dc.identifier.issn1932-6203
dc.identifier.doihttp://dx.doi.org/10.34944/dspace/5457
dc.identifier.other22242152 (pubmed)
dc.identifier.urihttp://hdl.handle.net/20.500.12613/5475
dc.description.abstractIntroduction: Compounds exhibiting low non-specific intracellular binding or non-stickiness are concomitant with rapid clearing and in high demand for live-cell imaging assays because they allow for intracellular receptor localization with a high signal/noise ratio. The non-stickiness property is particularly important for imaging intracellular receptors due to the equilibria involved. Method: Three mammalian cell lines with diverse genetic backgrounds were used to screen a combinatorial fluorescence library via high throughput live cell microscopy for potential ligands with high in- and out-flux properties. The binding properties of ligands identified from the first screen were subsequently validated on plant root hair. A correlative analysis was then performed between each ligand and its corresponding physiochemical and structural properties. Results: The non-stickiness property of each ligand was quantified as a function of the temporal uptake and retention on a cell-by-cell basis. Our data shows that (i) mammalian systems can serve as a pre-screening tool for complex plant species that are not amenable to high-throughput imaging; (ii) retention and spatial localization of chemical compounds vary within and between each cell line; and (iii) the structural similarities of compounds can infer their non-specific binding properties. Conclusion: We have validated a protocol for identifying chemical compounds with non-specific binding properties that is testable across diverse species. Further analysis reveals an overlap between the non-stickiness property and the structural similarity of compounds. The net result is a more robust screening assay for identifying desirable ligands that can be used to monitor intracellular localization. Several new applications of the screening protocol and results are also presented. © 2012 Nath et al.
dc.format.extente28802-e28802
dc.language.isoen
dc.relation.haspartPLoS ONE
dc.relation.isreferencedbyPublic Library of Science (PLoS)
dc.rightsCC BY
dc.subjectAnimals
dc.subjectArabidopsis
dc.subjectCell Line
dc.subjectCell Survival
dc.subjectCombinatorial Chemistry Techniques
dc.subjectFluorescent Dyes
dc.subjectHumans
dc.subjectLigands
dc.subjectMice
dc.subjectMicroscopy
dc.subjectSmall Molecule Libraries
dc.titleIdentification of fluorescent compounds with non-specific binding property via high throughput live cell microscopy
dc.typeArticle
dc.type.genreJournal Article
dc.relation.doi10.1371/journal.pone.0028802
dc.ada.noteFor Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu
dc.date.updated2021-01-31T22:39:39Z
refterms.dateFOA2021-01-31T22:39:44Z


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