Abstract
This chapter presents the microassay procedure for nicotinamide adenine dinucleotide (NAD) + -dependent 15-hydroxyprostaglandin dehydrogenase. The assay procedure is based upon the determination of the amount of substrate prostaglandin (PG) E 1 lost during a reaction with NAD + -dependent 15-hydroxyprostaglandin dehydrogenase (NAD + - PGDH) in the tissue sample. The reaction is carried out in a small reaction volume at 37° for the specified time, then terminated. The amount of unreacted PGE 1 is measured in a coupled enzyme system. The initial reaction may be carried out on nanogram quantities of freeze-dried tissue in a reaction volume of 0.1 μL suspended in oil. Conversely, the microassay may be applied to larger sample sizes in a 1-ml reaction volume. The quantitative histochemical assay consists of seven steps. The procedure is described for NAD + -PGDH in kidney.
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