A sensitive enzyme-release assay for natural cytotoxicity is described. The kinetic determination of the amount of the enzyme lactate dehydrogenase (LDH) released from lysed target cells was determined to provide a sensitive and precise measure of natural cytotoxicity when used in conjunction with appropriate controls and calculational methods. Values for the percentage of cytotoxicity or kinetic parameters determined by this method were identical, within experimental error, to values determined in parallel 51Cr release assays. Moreover, it was found that the spontaneous release of LDH from the target cells tested was considerably lower than the spontaneous release of 51Cr. This enzyme-release cytotoxicity assay is convenient, inexpensive, and precise, and should be applicable to the study of other cytotoxicity reactions, including antibody-dependent and T-cell mediated reactions.