A luminescence-based test for determining ornithine decarboxylase activity

Yongchun Wang, Uriel Bachrach

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

A sensitive chemiluminescence-based method for the assay of ornithine decarboxylase (ODC) has been developed. This method, which permits the detection of putrescine (the product of ODC) at a picomolar range, can be used to determine ODC activity in cellular extracts. Extracts are incubated with ornithine and spotted onto p81 phosphocellulose paper strips. After drying, the papers are washed with ammonium hydroxide to remove contaminants, which may interfere with the assay. Putrescine is next eluted from the paper by shaking in an elution buffer containing magnesium sulfate. Partially purified hog kidney diamine oxidase is then used to oxidize putrescine in the eluate. The hydrogen peroxide formed during the oxidation is determined by chemiluminescence using luminol and peroxidase. This simple analytical method has the sensitivity of conventional assays based on the use of radioactive ornithine. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)299-302
Number of pages4
JournalAnalytical biochemistry
Volume287
Issue number2
DOIs
StatePublished - Dec 15 2000
Externally publishedYes

Keywords

  • Assay
  • Chemiluminescence
  • Diamine oxidase
  • Ornithine decarboxylase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'A luminescence-based test for determining ornithine decarboxylase activity'. Together they form a unique fingerprint.

Cite this