Poor anticoagulant response to activated protein C (APC), present in 20-60% of thrombophilic patients, is most often caused by abnormal factor Va due to the mutation of Arg506 to Gln, and DNA sequencing confirms this finding. At Scripps Reference Laboratory (SRL), we have validated an in-house assay to detect APC resistance. A study of 80 normal subjects (40 males and 40 females, 21-60 years old) showed that adult males and females have statistically significant differences in their anticoagulant response to APC. Furthermore, APC response is increased in older individuals. APC responses of the same 80 normal samples determined using the SRL assay were compared to a commercial kit (Chromogenix, Coatest). Although both procedures are similar, the SRL assay gave a greater difference between male and female normal ranges. Eight of 18 (44%) thrombophilic patients were identified as APC-resistant. Samples from individuals having the factor V Arg506Gln mutation yielded low responses to APC in both assays. Determination of APC resistance assists physicians in determining the cause of thrombotic disorders and may eventually help in predicting the risk of familial thrombosis.