Spectrophotometric assay for ornithine decarboxylase |
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| Authors: | That T. Ngo Kurt L. Brillhart Rowland H. Davis Raphael C. Wong Jack H. Bovaird Joseph J. Digangi Janet L. Ristow J. Lawrence Marsh Andrew P. H. Phan Howard M. Lenhoff |
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| Affiliation: | 1. Department of Food Science and Human Nutrition, College of Agriculture, Consumer and Environmental Sciences, University of Illinois, Urbana, IL, USA;2. Department of Food Science and Engineering, Faculty of Agricultural Engineering and Technology, University of Tehran, Karaj, Iran;3. School of Biology, College of Science, University of Tehran, Tehran, Iran;4. Nanobiomedicine Center of Excellence, Nanoscience and Nanotechnology Research Center, University of Tehran, Tehran, Iran;5. Center of Excellence for Application of Modern Technologies for Producing Functional Foods and Drinks, Iran;6. Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran;7. Department of Chemical and Bimolecular Engineering, School of Chemical Sciences, University of Illinois, Urbana, IL, USA;1. College of Chemical and Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418, China;2. Key Laboratory of Synthetic Chemistry of Natural Substances, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Ling Ling Road 345, Shanghai 200032, China |
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| Abstract: | A rapid and sensitive spectrophotometric assay for ornithine decarboxylase is described. It is based on the observation that the product of ornithine decarboxylase, putrescine, reacts with 2,4,6-trinitrobenzenesulfonic acid to give a colored product soluble in 1-pentanol whereas ornithine does not. The amount of putrescine produced by the enzyme was determined by measuring the absorbance of the 1-pentanol extract of the reaction mixture at 420 nm, and by comparing the results to those obtained by the trapping of 14CO2 and by HPLC assays. The three assays were found to be equivalent in sensitivity, with the spectrophotometric assay having the advantages of being relatively rapid, requiring only common laboratory equipment, and not requiring the use of radioactive isotopes. |
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| Keywords: | ornithine decarboxylase decarboxylases polyamines trinitrobenzene sulfonic acid spectrophotometry cancer diagnosis |
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