Sandwich enzyme immunoassay for ornithine decarboxylase.
A sensitive enzyme immunoassay (EIA) was developed for the determination of ornithine decarboxylase (ODC, EC 4.1.1.17), in the range of 0.02-10 ng, using an affinity-purified anti-ODC-Fab'-peroxidase conjugate. The amount of ODC protein was determined in crude extracts from the kidney of testosterone-treated mice, regenerating rat liver and human thyroid carcinoma, with purified mouse kidney or rat liver enzyme as standard. In all these tissues, similar activity/protein ratios were found for ODC: 1.2 x 10(6)-1.9 x 10(6) nmol CO2/h/mg of ODC protein, which were roughly equivalent to the final specific activity of purified enzymes. ODC inactivated by alpha- difluoro-methylornithine (DFMO) could also be assayed with this method similarly to active ODC protein. However, ODC-antizyme complex gave a somewhat lower value than free ODC protein.
Nishiyama
M
M
Department of Nutrition, Jikel University School of Medicine, Tokyo, Japan.
Matsufuji
S
S
Kanamoto
R
R
Murakami
Y
Y
Hayashi
S
S
D000818:Animals; D000518:Eflornithine / Q000494:pharmacology; D006801:Humans; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000187:drug effects / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008115:Liver Regeneration; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009955:Ornithine Decarboxylase / Q000032:analysis* / Q000276:immunology; D065108:Ornithine Decarboxylase Inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D013739:Testosterone / Q000494:pharmacology; D013964:Thyroid Neoplasms / Q000201:enzymology
D065108:Ornithine Decarboxylase Inhibitors; D013739:Testosterone; D009955:Ornithine Decarboxylase; D000518:Eflornithine