Intact epidermal cell assay for ornithine decarboxylase activity
dc.contributor.author | Duell, Elizabeth A. | en_US |
dc.contributor.author | Roseeuw, Diane I. | en_US |
dc.contributor.author | Marcelo, Cynthia L. | en_US |
dc.date.accessioned | 2006-09-11T18:44:31Z | |
dc.date.available | 2006-09-11T18:44:31Z | |
dc.date.issued | 1982-03 | en_US |
dc.identifier.citation | Roseeuw, Diane I.; Duell, Elizabeth A.; Marcelo, Cynthia L.; (1982). "Intact epidermal cell assay for ornithine decarboxylase activity." Archives of Dermatological Research 273 (1-2): 137-148. <http://hdl.handle.net/2027.42/47233> | en_US |
dc.identifier.issn | 1432-069X | en_US |
dc.identifier.issn | 0340-3696 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/47233 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=7184470&dopt=citation | en_US |
dc.description.abstract | A procedure measuring the ornithine decarboxylase (ODC) activity and polyamine formation of intact neonatal mouse epidermal cells in culture has been developed and tested. Basal cells prepared from neonatal mouse epidermis were plated on round 15-mm Lux coverslips, placed in Costar 24 well culture clusters and grown at 32°C in M-199 + 13% fetal bovine serum. Before assay the cells were rendered permeable to ornithine 14 C and ODC inhibitors using the buffer described by Berger et al. [3]. The slides, covered with adhering cell layers, were then placed in vials, covered with assay buffer and assayed intact for ODC activity. The ODC reaction was terminated by addition of citric acid to the buffer and the amount of 14 CO 2 released was determined by scintillation counting of a center well filled with trapping agent. The baseline ODC activity of the intact cells was 500–1,000 pmol 14 CO 2 /mg protein/45 min. The validity of this ODC assay procedure using intact neonatal mouse keratinocytes was tested by use of three specific ODC inhibitors and by measuring the formation of polyamines from uniform labeled ornithine. The results indicated that authentic ODC activity was measured and preserved in this intact neonatal mouse epidermal cell assay. This technique holds promise for future studies of epidermal cell regulation of ODC and polyamine synthesis and studies of the multiple ornithine metabolites and conjugates formed, using a highly manipulable in vitro system. | en_US |
dc.format.extent | 1198648 bytes | |
dc.format.extent | 3115 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Springer-Verlag; Springer-Verlag GmbH & Co KG | en_US |
dc.subject.other | Epidermal Cell Culture | en_US |
dc.subject.other | Primary Keratinocytes | en_US |
dc.subject.other | Dermatology | en_US |
dc.subject.other | Intact Epidermal Cell ODC Activity | en_US |
dc.subject.other | Medicine & Public Health | en_US |
dc.subject.other | Polyamine Metabolites and Conjugates | en_US |
dc.title | Intact epidermal cell assay for ornithine decarboxylase activity | en_US |
dc.type | Article | en_US |
dc.subject.hlbsecondlevel | Dermatology | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Dermatology, University of Michigan Medical School, 48109, Ann Arbor, MI, USA | en_US |
dc.contributor.affiliationum | Department of Dermatology, University of Michigan Medical School, 48109, Ann Arbor, MI, USA | en_US |
dc.contributor.affiliationother | Department of Dermatology, Adjunkt-Kliniekhoofd, Vrye Universiteit Brussel, AZ-VUB, Laarbeeklaan 101, B-1090, Brussels, Belgium | en_US |
dc.contributor.affiliationumcampus | Ann Arbor | en_US |
dc.identifier.pmid | 7184470 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/47233/1/403_2004_Article_BF00509038.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1007/BF00509038 | en_US |
dc.identifier.source | Archives of Dermatological Research | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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