Actin, labelled with the fluorescent dye N-(3-pyrenyl)maleimide, was diluted below its critical concentration and depolymerization was followed by measuring the declining fluorescence intensity. The time courses of depolymerization were fitted to a sum of three exponentials. In most cases there was a fast initial phase followed by one or three slower ones. Increasing MgCl2 concentration slowed down depolymerization velocity, as did substitution of Tris-maleate buffer by phosphate buffer. Older F-actin preparations depolymerized more slowly than younger ones. Phalloidin strongly decreased depolymerization velocity even after sonication. In the presence of cytochalasin B depolymerization was more uniformly exponential than in the absence of cytochalasin B; overall depolymerization velocity was decreased by cytochalasin B. The results are discussed on the assumption that depolymerization kinetics reflect the length distribution of actin filaments during depolymerization.