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Detection of LRV with a monocolonal anti-dsRNA (J2) antibody by immunofluorescence microscopy.

By Haroun Zangger (289611), Catherine Ronet (289613), Chantal Desponds (289615), F. Matthew Kuhlmann (256084), John Robinson (289616), Mary-Anne Hartley (289617), Florence Prevel (289618), Patrik Castiglioni (289619), Francine Pratlong (50534), Patrick Bastien (68928), Norbert Müller (244550), Laurent Parmentier (289620), Nancy Gore Saravia (169839), Stephen M. Beverley (174720) and Nicolas Fasel (209333)


<p><b>A.</b> Reference strain analysis (protocol A, see “Material and methods”). Green: dsRNA (J2 Ab). Blue: DAPI (standardized exposure time in all images). <b>B.</b> Phase and immunofluorescent images of <i>Lg</i> M4147 LRV<sup>high</sup> or LRV<sup>neg</sup> cells were obtained in the presence or absence of J2 antibody (protocol B). <b>C.</b> Quantitative immunofluorescence (protocol B). The fluorescent intensity per cell was assessed using Image J software on <i>Lg</i> M4147 LRV<sup>high</sup> or LRV<sup>neg</sup> cells following IFM with the J2 antibody. Cells from phase images were identified and the fluorescent intensity average over the area of the cell was recorded. 108–160 cells from 2 distinct fields were measured, and histogram plots were made using Excel software. LRV<sup>high</sup>, no primary antibody (▪, dashed line); LRV<sup>high</sup> with J2 (▪, solid line); LRV<sup>neg</sup>, no primary antibody (•, dashed line); LRV<sup>neg</sup> with J2 (•, solid line).</p

Topics: Microbiology, lrv, monocolonal, anti-dsrna, antibody, immunofluorescence
Year: 2013
DOI identifier: 10.1371/journal.pntd.0002006.g003
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Provided by: FigShare
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