Simplified procedure for preparation of35S-labeled brain sulfatide |
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Authors: | Arvan L. Fluharty Marsha L. Davis Hayato Kihara Gene Kritchevsky |
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Affiliation: | 1. Neuropsychiatric Institute-Pacific State Hospital Research Group Pomona, 91766, California 2. Division of Neurosciences, City of Hope National Medical Center, 91010, Duarte, California
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Abstract: | A simplified procedure for the preparation of35S-labeled brain cerebroside sulfates has been developed. The labeled sulfatides are synthesized in vivo after intracerebral administration of inorganic35S-sulfate in developing rats. The animals are sacrificed three days later and brain homogenates extracted with chloroform-methanol. The extract is subjected to mild alkaline treatment and washed with water. The organic phase is chromatographed on triethylaminoethyl-cellulose from which sulfatides are eluted with chloroform-methanol containing potassium acetate. Radioactive fractions are pooled, concentrated, and potassium acetate removed by dialysis against water. Alternatively, salts are removed by passing radioactive fractions through Sephadex G-25. After evaporating to near dryness, the radioactive cerebroside sulfates are dissolved in a small volume of chloroform-methanol and stored at −20 C. The35S-sulfatides are essentially free of lipid contaminants, and 97% of the radio-activity corresponds with sulfatides on chromatographic analysis. |
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