Glucocorticoids are involved in the regulation of metabolic processes of carbohydrate, lipid and protein, and have an almost systemic effect, e.g., blood pressure, immune system, nervous system and stress. Glucocorticoid levels in the blood circulation are controlled by a negative feedback mechanism of the hypothalamic-pituitary-adrenal axis. On the other hand, in peripheral tissues, intracellular levels of glucocorticoid are regulated by two isozymes, 11b-hydroxysteroid dehydrogenase type 1 and type 2 (11b-HSD-1 and 11b-HSD-2). They catalyze the interconversion of biologically inert 11-keto-steroid (11-dehydrocorticosterone or cortisone) and active 11-oxo-glucocortiocoids (corticosterone or cortisol). 11b-HSD-1, which exists in the liver, adipose tissue, brain, lung and gonads, reduces the carboxyl group at the C-11 position to a hydroxyl group to increase active intracellular glucocorticoid levels. Conversely, 11b-HSD-2 oxidizes the hydroxyl to a carboxyl group at the C-11 position in the distal nephron, colon and sweat glands to protect the mineralocorticoid receptor from binding abundant glucocorticoid in the circulation. [1][2][3][4] 11b-HSDs modulate the intracellular levels of active glucocorticoid, which exerts physiological effects, as described above. Thus, the disruption of their enzymatic regulation in peripheral tissues induces various diseases, for example, increased 11b-HSD-1 activity in the adipose tissues and liver is linked to metabolic syndrome, including obesity, insulin resistance and hypertension, 1,2,5,6) and decreased or deficient 11b-HSD-2 in the renal tubules causes hypertension and apparent mineralocorticoid excess.7-9) Therefore, it is clinically important to evaluate the activity of 11b-HSD-1 and 11b-HSD-2 in vivo; however, it is difficult to measure these activities directly in the tissues in vivo. One alternative is to measure the ratios of serum concentrations of cortisol and cortisone after the oral administration of cortisone acetate under the suppression of endogenous cortisol by dexamethasone.10-12) In mice and rats, which are the most conventional and standard experimental animals, cortisol and cortisone in humans correspond to corticosterone and 11-dehydrocorticosterone, respectively. Therefore, the determination of serum corticosterone and 11-dehydrocorticosterone levels is required to measure 11b-HSD activity in these animals 13) ; however, no convenient and useful enzyme immunoassay for 11-dehydrocorticosterone is available to date. 14,15) In this paper, we described ELISA for serum 11-dehydrocorticosterone and also investigated the developmental changes of serum 11-dehydrocorticosterone levels in rats and mice, and using the proposed ELISA, chronological changes of serum 11-dehydrocorticosterone levels after 11-dehydrocorticosterone administration to rats have been determined. Serum corticosterone levels were also measured in these rats. The results indicate that the proposed ELISA for 11-dehydrocorticosterone is useful for measuring 11b-HSD activities in combination wit...