Various endotoxins and the ether extracts of grampositive bacteria were measured immunologically by radioimmunoassay and also biologically by the Limulus test. The minimum amount of endotoxin detectable with the Limulus test was in the range from 1 ng/ml to 1 ..mu..g/ml, with the lysate of sensitivity, 100 ng ml (E. coli 0111: B4(B) lipopolysaccharide). On the other hand, by the radioimmunoassay they were estimated in the range of 0.3 to 10 times of dry weight. Endotoxin-like activity was detected in the ether extracts of grampositive bacteria at a minimum concentration between 1 ..mu..g/ml and 100 ..mu..g/ml with the Limulus test. However, most of them were estimated by the radioimmunoassay to be under 1/50 of dry weight. Various substances such as thrombin, thromboplastin, polynosinic-polycytidylic acid, polyadenylic-polyuridylic acid, carrageenan and human colonic mucosal antigen had cross reactivities of various degrees in the minimum concentration from 10 ..mu..g/ml to 10 mg/ml. Compounds such as thrombin and thromboplastin cross-reacting in the Limulus test were scarcely measured by the radioimmunoassay except for polynucleotides. From this study, it has become clear that the radioimmunoassay method is quite specific and accurate for quantitative measurements of endotoxin.