In situ detection of diamine oxidase activity using enhanced chemiluminescence.
In need of a simple and sensitive method for detection of diamine oxidase (EC 1.4.3.6) activity in connection with diamine oxidase purification from human placenta, we have developed an enhanced chemiluminescence method using putrescine as substrate and horseradish peroxidase and luminol for the detection of the H2O2 produced by diamine oxidase. The method allows direct detection of small amounts of diamine oxidase in serum samples after agarose gel electrophoresis and allows visualization of diamine oxidase activity in tissue sections. Employing this method we have detected diamine oxidase in sera from cow, horse, monkey, rabbit, and pregnant women. On tissue sections from term human placenta diamine oxidase activity was exclusively localized to the maternal side and was concentrated in vessels and fibrinoid areas.
Bruun
L
L
Department of Autoimmunology, Statens Seruminstitut, Copenhagen, Denmark.
Houen
G
G
D006631:Amine Oxidase (Copper-Containing) / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D000818:Animals; D002417:Cattle; D002522:Chlorocebus aethiops; D005260:Female; D006651:Histocytochemistry; D006735:Horseradish Peroxidase; D006736:Horses; D006801:Humans; D008163:Luminescent Measurements; D008165:Luminol; D010920:Placenta / Q000033:anatomy & histology / Q000201:enzymology; D011247:Pregnancy; D011700:Putrescine; D011817:Rabbits; D013379:Substrate Specificity
D008165:Luminol; D006735:Horseradish Peroxidase; D006631:Amine Oxidase (Copper-Containing); D011700:Putrescine