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Cultivation of S. aureus in the presence of 0.03 IU benzyl-penicillin stimulated the formation of a haemolysin which was suggested to be alpha-lysin. Its molecular weight was estimated to be 30000 by molecular sieve chromatography. It was mainly active on rabbit erythrocytes and was neutralized by alphalysin antibody. Furthermore it gave a reaction of identity in comparative immunoelectrophoresis with an are which was shown to represent alpha-lysin.