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作者首先使用免疫印迹和免疫沉淀法对抗APC羧基末端抗体进行评价,然后用这些抗体确定被截断的APC在体内是否与野生型APC相联。与用SW480所得结果相反,当检测从FAP病人衍生的淋巴母细胞样细胞株时,被截断的APC存在于免疫沉淀物中。此外,在细胞溶解产物中,截断的APC明显比野生型APC多,但在这些溶解产物的免疫沉淀中,突变APC的量等于或小于野生型APC。结果表明,这种被截断的蛋白质在体内与野生型APC相聚,且只有这种与野生型APC相连的突变APC可被免疫沉淀。当SW480溶解产物与由表达完整
The authors first evaluated anti-APC carboxy-terminal antibodies using immunoblotting and immunoprecipitation and then used these antibodies to determine whether truncated APCs are associated with wild-type APCs in vivo. In contrast to the results obtained with SW480, truncated APCs are present in immunoprecipitates when detecting lymphoblastoid cell lines derived from FAP patients. In addition, truncated APCs were significantly more truncated in cell lysates than wild-type APCs, but the amount of mutant APCs was equal to or less than that of wild-type APCs in immunoprecipitates of these lysates. The results show that this truncated protein is aggregated with the wild-type APC in vivo, and only this mutant APC linked to the wild-type APC can be immunoprecipitated. When SW480 lysate is expressed by intact