The result of my study may be summarized as follows:_??_It has made clear the fact that, hemoglobino-precipitin can be produced by immunizing with globin alone, this being a constituent of hemoglobin used as an antigen in a similar manner as if hemoglobin has been used. To summarize the results they are as follows:_??_
1. Globin, obtained by any method whatever, always produces a precipitin for itself and simultaneously it also produces hemoglobino-precipitin, though with a difference in its extent; in other words, the globin possesses both precipitinogenic property for globin at same time as for hemoglobin.
2. The globin, however, if once precipitated with ammonia, decreases its precipitinogenic property for hemoglobin, yetis still rich in precipitinogenic property for globin, but on the contrary, Schulz's globin showed a marked precipitinogenic property for hemoglobin excelling that for globin.
3. Since, without exception, globin showed presence of both precipitinogens mixed, there being consequently presence of precipitins mixed for the corresponding precipitinogen in anti-serum, these can be separated by application of the absorption method rom these antisera.
4. Although, the globin, which has been secured from decaying globin, produced precipitin, yet almost lost its species-specificity, all the other globins showed very distinct species-specificity to the antisera applied.
5. The phosphatide, which has been extracted with alcohol from hemoglobin and red cells, does not exert any influence on species-specificity, even though immunized by addition to globin, also it showed no trace of a relation to formation of hemoglobino-precipitinogen, save that it is likely to activate and accelerate the formation of hemoglobino-precipitin.
6. Use of alcohol in the purification process of globin markedly interferes the precipitinogenic property of hemoglobin and this is, probably, largely due to its chemical adulteration. Holden and Freeman's native globin is found to be most rich in hemoglobinoprecipitinogenic property.
7. This native globin, when treated with alcohol, can be separated to supernatant fluid and precipitate and it showed itself to be almost devoid of phosphorus matter in the precipitate. Further, althoughitlacks precipitinogenic property for hemoglobin, yet, itproved to possess precipitinogenic property for globin and its speciesspecificity. This noteworthy fact seems to indicate that, the precipitinogenic property for hemoglobin is largely related with a presence of ordinary globin in combination with phosphorus (probably phosphatide). However, the species-specificity is not influenced by this factor.
8. Through the results of my study above described, it is clear that, the precipitinogenic property of hemoglobin is based on the substance, globin, which is a component of hemoglobin, as has already been stated by Ishikawa and Sakurabayashi.
The result of my present study, when discussed with references to the work and observation of other workers, shows, as has already been stated in the introductory part of this paper, that, although the fact of the antigenic property of globin cannot be doubted at present, yet, the capability of production of precipitin for hemoglobin has been restricted solely to hemoglobin, the role of globin being neglected and even denied by many authors, while the view of Ishikawa and Sakurabayashi who claimed in 1925 that globin, instead of hemoglobin, is the base of antigen in the formation of hemoglobine-precipitin, was not paid much attention by many workers except by Hektoen and Schulhof a year later.
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