【摘要】 目的:改进Western blotting转膜方法,提高低丰度蛋白转膜效率。方法:在Western blotting转膜过程中,采用不同孔径的PVDF膜、不同的转膜时间及不同的甲醇含量进行转膜。转膜后,考马斯亮蓝染色观察PAGE胶上蛋白残留情况,ECL发光法检测FGFR1曝光强度。结果:通过改变膜孔径大小和转膜时间所得实验结果可见,孔径为0.22μm的PVDF膜转移2h的转膜效率更高。通过改变转膜液中甲醇含量所得实验结果可见,甲醇含量为10%转膜效率更高。结论:本实验对Western blotting转膜过程中低丰度蛋白的转膜方法进行了成功改进,改进后能提高蛋白显影强度,具有广泛的应用前景,是一种方便可行的操作方法。更多还原

【Abstract】 Objective: To optimize the condition of transfer process for raising the membrane-transferring efficiency of low abundance protein in Western blotting. Methods: Different pore diameter PVDF membrane,different transfer time and different methyl alcohol concentration were tested in the process of membrane-transfer;then the efficiency was compared through coomassie brilliant blue staining and ECL. Results: The optimun experimental condition for low abundance pratein is the PVDF membrane with 0.22 μm pore diameter, the transfer time for 2 hours and the methanol concentration at 10%. Conclusion: The improved transfer process in Western blotting can strengthen the development of low abundunce protein. It has broad application prospects.更多还原