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Spectrophotometric assay for ornithine decarboxylase
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  
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.
Keywords:ornithine decarboxylase  decarboxylases  polyamines  trinitrobenzene sulfonic acid  spectrophotometry  cancer diagnosis
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