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己有的用于测定脑内ACh的许多方法中,存在专一性和重复性较差、测定前需抽提或制备成衍生物等缺点。木文采用了简便灵敏的放射免疫分析,测定脑提取液中微量的ACh。免疫原制备及免疫:在1g氯化胆碱和10ml于吡啶的混合物内加入4g戊二酸酐,加10mlN、N-二甲基甲酰胺使溶解,搅拌使渐形成白色沉淀。室温下搅拌过夜,然后过滤,用乙酸乙酯冲洗,加丙酮-甲醇使析出半戊二酸胆碱。将100mg小牛血清白蛋白溶于10ml0.2M的磷酸缓冲液中(pH6.0),边搅边加20mg半戊二酸胆碱和100mg1-乙基-3-(3-二甲胺基
Some of the methods that have been used to measure ACh in the brain have disadvantages such as poor specificity and poor reproducibility, extraction or preparation of a derivative before measurement. Mu Wen used a simple and sensitive radioimmunoassay, determination of trace amount of brain extract ACh. Preparation and Immunization of Immunogen: 4 g of glutaric anhydride was added to 1 g of choline chloride and 10 ml of pyridine, and 10 ml of N, N-dimethylformamide was added to dissolve and stir to form a white precipitate. Stir overnight at room temperature, then filter, rinse with ethyl acetate, add acetone-methanol to precipitate choline semiglutarate. 100 mg of bovine serum albumin was dissolved in 10 ml of 0.2 M phosphate buffer (pH 6.0), and 20 mg of choline pentachymate and 100 mg of 1-ethyl-3- (3-dimethylamino