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Detecting intracellular translocation of native proteins quantitatively at the single cell level

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dc.contributorVirginia Tech. School of Biomedical Engineering and Sciencesen
dc.contributorVirginia Tech. Department of Biological Sciencesen
dc.contributorVirginia Tech. Department of Chemical Engineeringen
dc.contributor.authorCao, Zhenningen
dc.contributor.authorGeng, Shuoen
dc.contributor.authorLi, Liwuen
dc.contributor.authorLu, Changen
dc.contributor.departmentSchool of Biomedical Engineering and Sciencesen
dc.date.accessed2015-04-16en
dc.date.accessioned2015-04-20T22:22:11Zen
dc.date.available2015-04-20T22:22:11Zen
dc.date.issued2014-04-07en
dc.description.abstractThe intracellular localization and movement (i.e. translocation) of proteins are critically correlated with the functions and activation states of these proteins. Simple and accessible detection methods that can rapidly screen a large cell population with single cell resolution have been seriously lacking. In this report, we demonstrate a simple protocol for detecting translocation of native proteins using a common flow cytometer which detects fluorescence intensity without imaging. We sequentially conducted chemical release of cytosolic proteins and fluorescence immunostaining of a targeted protein. The detected fluorescence intensity of cells was shown to be quantitatively correlated to the cytosolic/nuclear localization of the protein. We used our approach to detect the translocation of native NF-_B (an important transcription factor) at its native expression level and examine the temporal dynamics in the process. The incorporation of fluorescence immunostaining makes our approach compatible with the analysis of cell samples from lab animals and patients. Our method will dramatically lower the technological hurdle for studying subcellular localization of proteins.en
dc.description.notesSupplementary information is included in a separate fileen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationCao, Z., Geng, S., Li, L., & Lu, C. (2014). Detecting intracellular translocation of native proteins quantitatively at the single cell level. Chemical Science, 5(6), 2530-2535. doi: 10.1039/C4SC00578Cen
dc.identifier.doihttps://doi.org/10.1039/C4SC00578Cen
dc.identifier.issn2041-6520en
dc.identifier.urihttp://hdl.handle.net/10919/51724en
dc.identifier.urlhttp://pubs.rsc.org/en/content/articlelanding/2014/sc/c4sc00578cen
dc.language.isoenen
dc.publisherThe Royal Society of Chemistryen
dc.rightsCreative Commons Attribution-NonCommercial 3.0 Unporteden
dc.rights.urihttp://creativecommons.org/licenses/by-nc/3.0/en
dc.subjectFluorescence microscopyen
dc.subjectFlow cytometry analysisen
dc.subjectStandard fluorescence immunostainingen
dc.subjectWestern blottingen
dc.titleDetecting intracellular translocation of native proteins quantitatively at the single cell levelen
dc.title.serialChemical Scienceen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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