蛋白质酪氨酸磷酸化-免疫沉淀及Western印迹两种方法的比较

Tyrosine phosphorylation of proteins in murine thymocytes-a comparison between immunoprecipitation and Western blotting

为了深入了解在细胞信息传递中蛋白质酪氨酸磷酸化研究的较佳方法，采用免疫沉淀法及Ｗｅｓｔｅｒｎ印迹法对电离辐射诱导的小鼠胸腺细胞蛋白质磷酸化进行了比较分析。免疫沉淀结果表明，照射后１７５００、２２０００、２８０００、３２０００、４００００、４３０００及５３０００蛋白分子发生酪氨酸磷酸化。Ｗｅｓｔｅｒｎ印迹结果表明，照射后２８０００、３２０００蛋白分子发生酪氨酸磷酸化。提示在蛋白质酪氨酸磷酸化的研究中，免疫沉淀法优于Ｗｅｓｔｅｒｎ印迹法。

A comparison between the two methods was performed for tyrosine phosphorylation of proteins in murine thymocytes exposed to low dose radiation using antiphosphotyrosine McAb.It was shown that tyrosine phosphorylation of 17.5 kD,22 kD,28 kD,32 kD,40 kD,43 kD and 53 kD protein molecules labelled metabolically with H332PO4 appeared in the thymocytes after radiation by immunoprecipitation technique and tyrosine phosphorylation of 28 kD,32 kD protein molecules appeared by western blotting.The results suggest that immunoprecipitation is superior to Western blotting in detection of protein tyrosine phosphorylation.