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Assay of prolyl hydroxylase in cultured fibroblast monolayers

โœ Scribed by Joel S. Trupin; James D. Russell; Shirley B. Russell

Publisher
Elsevier Science
Year
1982
Tongue
English
Weight
364 KB
Volume
121
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

Prolyl hydroxylase may be assayed in cultured cell monolayers after a cold acetone treatment or air-drying. Enzyme activity of dried cells is stable when cell monolayers are stored frozen and desiccated, thus permitting simultaneous assay of cells harvested at different times, e.g., on different days of a culture cycle. Uniformity of cell monolayers permits use of separate dishes for assay of protein or DNA to determine specific enzyme activity. This procedure greatly facilitates harvesting and assay of cells for a variety of biochemical measurements,

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A procedure for the direct solubilization of fibroblast monolayers allowing precise determination of DNA and protein content has been developed. This simple procedure is applicable to the analysis of these macromolecules in fibroblast cultures containing small numbers of cells (<l x 105).