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DURING the investigation of a hmolytic transfusion reaction it was observed that the patient's serum contained an unusual antibody; tests against a large number of human bloods showed that the factor thus defined was one which had not previously been recognised. A brief account of these observations has already been published (Cutbush, Mollison and Parkin, 1950). The purpose of the present paper is to give a fuller account of the investigations and to present some additional observations. CASE HISTORYThe patient (R. D.), aged 43, was a sufferer from hmophilia.Signs of the disease were first manifest in infancy and there had since been many attacks of spontaneous bleeding. Up to July the patient had had only three blood transfusions. The first was given in 1928, when persistent bleeding followed the extraction of a tooth. This transfusion was followed by a rigor. The patient received his second and third transfusions in 1936 : both were followed by rigors but no further details are available. One week before the first of these two transfusions the patient received an intramuscular injection of blood.The patient then remained in moderately good health until 7th July 1949, when he was admitted to hospital with spontaneous bruising and bleeding. On 11th July, the patient was in need of a transfusion; he was found to be group 0 Rh negative and the blood of 3 group 0 Rh negative donors was tested against his serum as follows : The cells of each donor, suspended in 20 per cent. albumin, were mixed with an equal volume of the patient's serum ; the mixtures were left at 370 C. for an hour and the sediments were then examined microscopically. No trace of agglutination was observed in any of the three mixtures. The transfusion was started and, although the patient developed a rigor during its course, the blood from all three donors was eventually given.On the following day the patient became jaundiced and further investigations were therefore undertaken.It was now found that using the indirect Coombs test, the patient's serum reacted with the blood of the first and third donors, though not with the blood of the second donor. The fate of the cells from the third donor could be followed, since the donor belonged to 383