The commercially available trisodium salt of adenosine 5'-tetraphosphate (ATetraP) (Sigma) was found to be contaminated with ATP, ADP and AMP, and therefore unsuitable for use in platelet studies. 2. The more stable barium salt of ATetraP was converted to the ammonium salt and found to be chromatographically homogeneous. This sample was tested for its influence on sheep blood platelets in citrated-rich plasma by the photometric method. 3. The ammonium salt of ATetraP (5-105 mumol.1(-1)) induced platelet aggregation which showed no tendency towards disaggregation. 4. The log dose-response lines for ATetraP and for adenosine diphosphate were parallel. On a molar basis, the tetraphosphate and only 1.5% of the aggregating activity of ADP. 5. The initial rate of aggregation induced by the tetraphosphate was inhibited by adenosine 5'-monophosphate analogues which are selective ADP-antagonists. These compounds also dispersed aggregates produced by ATetraP. 6. Platelets made refractory to ADP were also refractory to ATetraP. 7. Like ADP, ATetraP induced the change in shape of rabbit platelets and in this respect had only 3.4% the activity of ADP. 8. It is concluded that ATetraP per se can induced platelet aggregation and platelet shape change, and appears to exert its effect at the same site on the platelet surface as does ADP.