Semi-preparative purification of an endogenous ligand for brain serotonin-2 receptors by coil planet centrifuge counter-current chromatography.
A horizontal flow-through coil planet centrifuge equipped with a rotatory frame holding three sets of composite column assemblies was used for purification of an endogenous ligand (ketanserin binding inhibitor) for the [3H]-ketanserin (3H-KET) recognition site. The protein mixture containing the endogenous material was successfully resolved by using a two-phase solvent system consisting of 95% ethanol-31.5% ammonium sulphate (1:2). The active fractions on 3H-KET binding obtained after counter-current chromatography (CCC) were further purified through a C18 microBondapak reversed-phase high-pressure liquid chromatographic column. The introduction of this advanced CCC technique represents an important step in the application of CCC for the separation of polar proteins from protein mixtures.
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FIDIA-Georgetown Institute for the Neurosciences, Washington, DC 20007.
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D000645:Ammonium Sulfate; D000818:Animals; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002498:Centrifugation; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D000431:Ethanol; D007650:Ketanserin; D011869:Radioligand Assay; D051381:Rats; D011985:Receptors, Serotonin / Q000378:metabolism*; D012997:Solvents; D014316:Tritium
D011985:Receptors, Serotonin; D012997:Solvents; D014316:Tritium; D000431:Ethanol; D007650:Ketanserin; D000645:Ammonium Sulfate