We have developed and characterized a highly sensitive ELISA for soluble Fas (sFas) in the serum. The linearity of calibrator range was 0.06-2.00 micrograms/L and the detection limit was 0.01 microgram/L. The average within- and between-run CVs were 3.9% and 3.8%, respectively. The recovery of added sFas to serum was 93-118%. The effects of possible interferences (tryglyceride, hemoglobin, bilirubin) were negligible. We determined serum sFas in 155 healthy subjects, ages 20-69. The mean value of sFas in men (2.50 +/- 0.63 micrograms/L, n = 78) was significantly higher than that in women (2.01 +/- 0.53 micrograms/L, n = 77) (P < 0.001). Furthermore, there was a significant correlation between serum sFas concentration and age (men, r = 0.397, P < 0.001; women, r = 0.569, P < 0.001). Although the concentrations of sFas tended to increase with aging, it remains to be clarified how Fas-mediated apoptosis relates to aging.