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10.14814/phy2.14485 http://scihub22266oqcxt.onion/10.14814/phy2.14485 32592328!7319944!32592328     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=32592328&cmd=llinks): Failed to open stream: HTTP request failed! 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Erythropoietin production by the kidney and the liver in response to severe hypoxia evaluated by Western blotting with deglycosylation |pdf=|usr=}}{{32592328}} gab.com Text Erythropoietin production by the kidney and the liver in response to severe hypoxia evaluated by Western blotting with deglycosylation JO: Physiol Rep http://www.kidney.de/pget.php?&tw=1&pmid=32592328 #FOAvip The detection of erythropoietin (Epo) protein by Western blotting has required pre-purification of the sample. We developed a new Western blot method to detect plasma and urinary Epo using deglycosylation. Epo in urine and tissue, and erythropoiesis-stimulating agents (ESAs) in urine were directly detected by our Western blotting. Plasma Epo and ESAs were not detected by direct application but were detected by our Western blotting after deglycosylation. The broad bands of Epo and ESAs were shifted to 22 kDa by deglycosylation except for PEG-bound epoetin beta pegol. The 22 kDa band from an anemic patient's urine was confirmed by Liquid Chromatography/Mass Spectrometry (LC/MS) to contain human Epo. Severe hypoxia (7% O(2,) 4 hr) caused a 400-fold increase in deglycosylated Epo expression in rat kidneys, which is consistent with the increases in both Epo gene expression and plasma Epo concentration. Immunohistochemistry showed Epo expression in nephrons but not in interstitial cells under control conditions, and hypoxia increased Epo expression in interstitial cells but not in tubules. These data show that intrinsic Epo and all ESAs can be detected by Western blot either directly in urine or after deglycosylation in blood, and that the kidney but not the liver is the main site of Epo production in control and severe hypoxia. Our method will make the tests for Epo doping and detection easy. Twit Text FOAVip Erythropoietin production by the kidney and the liver in response to severe hypoxia evaluated by Western blotting with deglycosylation ????Physiol Rep http://www.kidney.de/pget.php?&tw=1&pmid=32592328 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32592328 #FOAvip English Wikipedia <ref>{{Cite pmid|32592328}}</ref> Erythropoietin production by the kidney and the liver in response to severe hypoxia evaluated by Western blotting with deglycosylation #MMPMID32592328 Yasuoka Y; Fukuyama T; Izumi Y; Nakayama Y; Inoue H; Yanagita K; Oshima T; Yamazaki T; Uematsu T; Kobayashi N; Shimada Y; Nagaba Y; Mukoyama M; Yamashita T; Sato Y; Sands JM; Kawahara K; Nonoguchi H Physiol Rep 2020[Jun]; 8 (12): e14485 PMID32592328show ga The detection of erythropoietin (Epo) protein by Western blotting has required pre-purification of the sample. We developed a new Western blot method to detect plasma and urinary Epo using deglycosylation. Epo in urine and tissue, and erythropoiesis-stimulating agents (ESAs) in urine were directly detected by our Western blotting. Plasma Epo and ESAs were not detected by direct application but were detected by our Western blotting after deglycosylation. The broad bands of Epo and ESAs were shifted to 22 kDa by deglycosylation except for PEG-bound epoetin beta pegol. The 22 kDa band from an anemic patient's urine was confirmed by Liquid Chromatography/Mass Spectrometry (LC/MS) to contain human Epo. Severe hypoxia (7% O(2,) 4 hr) caused a 400-fold increase in deglycosylated Epo expression in rat kidneys, which is consistent with the increases in both Epo gene expression and plasma Epo concentration. Immunohistochemistry showed Epo expression in nephrons but not in interstitial cells under control conditions, and hypoxia increased Epo expression in interstitial cells but not in tubules. These data show that intrinsic Epo and all ESAs can be detected by Western blot either directly in urine or after deglycosylation in blood, and that the kidney but not the liver is the main site of Epo production in control and severe hypoxia. Our method will make the tests for Epo doping and detection easy. |Anemia/blood/urine[MESH] |Animals[MESH] |Blotting, Western/methods[MESH] |Disease Models, Animal[MESH] |Erythropoietin/*biosynthesis/blood/urine[MESH] |Glycosylation[MESH] |Humans[MESH] |Hypoxia/blood/*metabolism/urine[MESH] |Kidney/*metabolism[MESH] |Liver/*metabolism[MESH] |Male[MESH] |Rats[MESH]Erythropoietin production by the kidney and the liver in response to s(epmc).pdf DeepDyve Pubget Overpricing