Freezing of serum samples at -30 degrees C without protective agents is the simplest and least expensive method of storage in serum banks. We investigated the stability of creatine kinase (CK) in human sera after freezer storage under such conditions for 24 h (n = 30) or for 2 or 4 weeks (n = 99). CK activity was measured in fresh sera and compared to matched thawed sera after freezer storage at the designated time intervals. The enzyme's median activity decreased significantly after 24 h, 2 weeks, and 4 weeks of freezer storage by 2.6, 5.9, and 8.3%, respectively (p < 0.0001, r = 0.99). Sex or high CK initial values had no significant effect on these results. We conclude that freezer storage of serum at -30 degrees C, even for short periods, causes a steady and significant decline in CK activity. These results should be taken into consideration when analyzing CK activity in frozen sera for research or clinical purposes.