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10.1371/journal.pone.0179217 http://scihub22266oqcxt.onion/10.1371/journal.pone.0179217 C5467901!5467901!28604827     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=28604827&cmd=llinks): Failed to open stream: HTTP request failed! 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A novel in vivo method to quantify slit diaphragm protein abundance in murine proteinuric kidney disease |pdf=|usr=}}{{28604827}} gab.com Text A novel in vivo method to quantify slit diaphragm protein abundance in murine proteinuric kidney disease JO: PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=28604827 #FOAvip Injury of the glomerular filter causes proteinuria by disrupting the sensitive interplay of the glomerular protein network. To date, studies of the expression and trafficking of glomerular proteins have been mostly limited to in vitro or histologic studies. Here, we report a novel in vivo biotinylation assay that allows the quantification of surface expression of glomerular proteins in mice. Kidneys were perfused in situ with biotin before harvest. Afterwards glomeruli were isolated and lyzed. The protein of interest was separated by immunoprecipitation and the amount of surface-expressed protein was quantified by Western blot analysis with streptavidin staining. As proof-of-concept, we examined the presence of nephrin in the slit diaphragm in two well-established murine models of proteinuric kidney disease: nephrotoxic nephritis and adriamycin nephropathy. In proteinuric animals, significantly less nephrin was detected in the slit diaphragm. When proteinuria decreased once again during the course of disease, the amount of surface nephrin returned to the baseline. Our present results suggest that our assay is a valuable tool to study the glomerular filter in proteinuric kidney diseases. Note that the assay is not limited to proteins expressed in the slit diaphragm, and all surface proteins that are accessible to biotin perfusion and immunoprecipitation qualify for this analysis. Twit Text FOAVip A novel in vivo method to quantify slit diaphragm protein abundance in murine proteinuric kidney disease ????PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=28604827 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28604827 #FOAvip English Wikipedia <ref>{{Cite pmid|28604827}}</ref> A novel in vivo method to quantify slit diaphragm protein abundance in murine proteinuric kidney disease #MMPMID28604827 Haase R; Potthoff SA; Meyer-Schwesinger C; Frosch C; Wiech T; Panzer U; Königshausen E; Stegbauer J; Sellin L; Rump LC; Quack I; Woznowski M PLoS One 2017[]; 12 (6): ä PMID28604827show ga Injury of the glomerular filter causes proteinuria by disrupting the sensitive interplay of the glomerular protein network. To date, studies of the expression and trafficking of glomerular proteins have been mostly limited to in vitro or histologic studies. Here, we report a novel in vivo biotinylation assay that allows the quantification of surface expression of glomerular proteins in mice. Kidneys were perfused in situ with biotin before harvest. Afterwards glomeruli were isolated and lyzed. The protein of interest was separated by immunoprecipitation and the amount of surface-expressed protein was quantified by Western blot analysis with streptavidin staining. As proof-of-concept, we examined the presence of nephrin in the slit diaphragm in two well-established murine models of proteinuric kidney disease: nephrotoxic nephritis and adriamycin nephropathy. In proteinuric animals, significantly less nephrin was detected in the slit diaphragm. When proteinuria decreased once again during the course of disease, the amount of surface nephrin returned to the baseline. Our present results suggest that our assay is a valuable tool to study the glomerular filter in proteinuric kidney diseases. Note that the assay is not limited to proteins expressed in the slit diaphragm, and all surface proteins that are accessible to biotin perfusion and immunoprecipitation qualify for this analysis. äA novel in vivo method to quantify slit diaphragm protein abundance in(epmc).pdf DeepDyve Pubget Overpricing