Quantification of O -GlcNAc protein modification in neutrophils by flow cytometry This article is a US government work and, as such, is in the public domain in the United States of America.
dc.contributor.author | Madsen-Bouterse, Sally A. | en_US |
dc.contributor.author | Xu, Yi | en_US |
dc.contributor.author | Petty, Howard R. | en_US |
dc.contributor.author | Romero, Roberto | en_US |
dc.date.accessioned | 2008-07-01T14:11:04Z | |
dc.date.available | 2009-07-06T16:34:52Z | en_US |
dc.date.issued | 2008-07 | en_US |
dc.identifier.citation | Madsen-Bouterse, Sally A.; Xu, Yi; Petty, Howard R.; Romero, Roberto (2008). "Quantification of O -GlcNAc protein modification in neutrophils by flow cytometry This article is a US government work and, as such, is in the public domain in the United States of America. ." Cytometry Part A 73A(7): 667-672. <http://hdl.handle.net/2027.42/60232> | en_US |
dc.identifier.issn | 1552-4922 | en_US |
dc.identifier.issn | 1552-4930 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/60232 | |
dc.description.abstract | Observations of intracellular O -linked Β- N -acetylglucosamine ( O -GlcNAc) protein modification are primarily performed by Western blot or immunofluorescence microscopy. The goal of this study was to develop a flow cytometric-based assay for O -GlcNAc signaling and thus provide a more quantitative and rapid method to facilitate clinical analyses. Isolated peripheral blood neutrophils were stimulated with fMLF after adherence to glass cover slips. Cells in suspension were treated with either fMLF or PMA. Unstimulated cells served as controls. Neutrophils were fixed with formaldehyde and permeabilized with cold methanol before intracellular O -GlcNAc staining. Cells on cover slips were analyzed by fluorescence microscopy, and suspension cell data were acquired by flow cytometry. O -GlcNAc protein modification was increased following neutrophil stimulation with either 100 nM fMLF or 10 nM PMA. Increases were detected following either treatment using both flow cytometry and fluorescence microscopy. The time necessary for the completion of staining, data acquisition, and analysis was considerably less using flow cytometry. In addition, flow cytometry allows for the analysis of a substantially greater number of cells. Neutrophil protein modifications by O -GlcNAc are rapidly detected using flow cytometry and provide information similar to that observed using fluorescence microscopy. Published 2008 Wiley-Liss, Inc. | en_US |
dc.format.extent | 573062 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.publisher | Wiley Subscription Services, Inc., A Wiley Company | en_US |
dc.subject.other | Life and Medical Sciences | en_US |
dc.subject.other | Cell & Developmental Biology | en_US |
dc.title | Quantification of O -GlcNAc protein modification in neutrophils by flow cytometry This article is a US government work and, as such, is in the public domain in the United States of America. | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Molecular, Cellular and Developmental Biology | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Ophthalmology and Visual Sciences, The University of Michigan Medical School, Ann Arbor, Michigan 48105 | en_US |
dc.contributor.affiliationother | Perinatology Research Branch, NICHD/NIH/DHHS, Bethesda, Maryland 20892 | en_US |
dc.contributor.affiliationother | Perinatology Research Branch, NICHD/NIH/DHHS, Bethesda, Maryland 20892 | en_US |
dc.contributor.affiliationother | Perinatology Research Branch, NICHD/NIH/DHHS, Bethesda, Maryland 20892 ; Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, Michigan 48201 ; Perinatology Research Branch, NICHD, NIH, DHHS, 3990 John R, 4th Floor, Detroit, MI 48201, USA | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/60232/1/20569_ftp.pdf | |
dc.identifier.doi | http://dx.doi.org/10.1002/cyto.a.20569 | en_US |
dc.identifier.source | Cytometry Part A | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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