Thin Layer Autoradiographic Electrophoresis of Tritium-Labeled Compounds in Buffered Photographic Gelatin
Strips of Kodirex-X ray film, immersed for a few minutes into the chosen buffer, then quickly blotted between two sheets of filter paper, have been used as a supporting medium for the electrophoretic separation of various radioactive ions. After the electrophoretic run, the film strip was dried and stored in the dark until developed. Autoradiographic electrophoresis has been carried out with sulfate, taurine, taurocholate-35S, tritium-labeled succinic acid, histidine, glutamic acid-14C, tritium-labeled valine, and methionine. The separation of glutamic acid, histidine -14C, and tritium -labeled valine has been achieved at pH 7.25 and 5.25 (20 to 30 volts per centimeter for 60 min) (Fig. 1). The use of glucose-14C has shown that the nonelectrophoretic migration due to electro-osmosis and evaporation is negligible at the two above pH values (Fig. 2). The separations are sharp, the diffusion is weak, and unless the specific activities are very high, radioactive ions have no time to leave any track as they move in the photographic gelatin.
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