“Studies on the Separation and the Extractionof Anterior Pituitary Hormone by Paper-electrophoresis”

Abstract

Studies on the apparatus of horizontal paper-electrophoresis and continuous paper-electrophoresis with cooler. (1st Report) a) Employing the paper-electrophoresis of natural cooling system, the experiment at temperatue higher than 25°C in summer was found to be difficult for obtaining the clear separating figures.b) Utilizing the cooler, even under the room temperature higher than 25°C., which keeps it lower than the normal temperature. Freezing mixture, if employed, make experiments possible by its effect of lowering temperature ten more degrees.c) The filter paper wrapped around by plate glass reduced its vaporization to half, and the buffer action to the temp. is increases. So the temp. in the cooler is able to be kept constant.d) The filter paper should always kept strained, especially in high temp. in summer, not to inflates itself. A glass rod was used in order to increase the strain.e) Employing the continuous paper-electrophoresis, there is no combustion by drying of the surface of the filter paper. Utilizing the cooler, however, the vaporization and the formation of water drop are reduced and approximately constant low temp. can be held not be influenced by the atmosphere.f) Very clear separating figures were obtained using the thick filter paper donated by Dr. K. Hanning of Max Planck Inst. Toyo No. 26 filter (made in Japan) was found to be inadequate.The electrophoresis of TSH products on the filter paper. (2nd Report) a) The Pretiron of Schering is separated into more than 2-3 fractions at pH. around 8 on electrophoresis.b) On the paper-electrophoresis of TSH substances, the staining Amidoblack 10B, Veronal or phosphatic acid buffer solution with ionic strength 0.05-0.1 and pH 8-8.5, are the most suitable. It is also convenient to set the starting point at the center.c) TSH activity is not effective at pH. below 7, so it is suitable on electrophoresis to use the buffer solution at pH. above 8-8.5.d) It was recognized that TSH can be separated and refined without reducing its activity on electrophoresis under the condition of up to 1000 volts per 40 cm. and 4.6 mA. per 10 cm.The separation and the refinement of TSH and the transmission of TSH substances in vivo on the continuos paper-electrophoresis. (3rd Report) a) TSH activity is effective only at pH. near 8, which showes that TSH is separated itself from gonadotropin in the mixture. That is, gonadotropin alone can be picked up.b) It was recognized that amoung three compounds of Pretiron separated at pH. near 8, the fraction at the center showed the strongest TSH activity and that the fration towards negative pole showed no activity. It was also recognized that there is the gonadotropic activity at the center.c) In the blood in vivo, TSH was recognized to transmit with gamma-globulin.