| 王文倩 魏 颖 王 宇 徐玉东 杨永清△ 尹磊淼.蛋白免疫印迹法检测小分子蛋白的实验条件优化研究[J].,2015,15(7):1230-1232 |
| 蛋白免疫印迹法检测小分子蛋白的实验条件优化研究 |
| Optimization Research on the Western Blot Experimental Conditions forDetecting Small Molecule Proteins* |
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| DOI： |
| 中文关键词: 免疫印迹法 Western blot 小分子蛋白 甲醇 化学发光剂 |
| 英文关键词: Immunoblotting Western blot Small molecule protein Methanol Chemiluminescence reagent |
| 基金项目:国家自然科学基金项目 (81 17334； 811 73332； 81202753； 81 473760)；上海市卫生系统优秀青年人才培养计划(XYQ2013081)；
上海市中医药事业发展三年行动计划重大研究项目 (ZYSNXD-CC-ZDYJ039) |
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| 中文摘要: |
| 目的： 蛋白免疫印迹法自 发明以来被广泛应用 于现代生物学研究中的蛋白 质定性和半定量分析。为了 提高蛋白免疫印迹法
的检测效率， 需针对不同 蛋白的特性调节相关的实验条件参数。 本文旨 在探讨免疫印迹法不同 参数对小分子蛋白 检测效果的影
响， 从而优化并获得最佳实验条件。 方法： 比较不同 转膜电压和时间、转移缓冲液甲醇含量、不同 化学发光剂对小分子蛋白的检测
效果。 结果： 选择 20 V、1 0 min 转膜电压和时间所获得的信号显著高于 1 0 V、 25 min 转膜条件，选择含 20 %甲醇转移缓冲液所获
得的信号显著高于无甲醇转移缓冲液，选择飞克级化学发光剂 所获得的信号显著高于纳克级化学发光剂。 结论： 选用 高电压、短
时间 组合，选择含 20 %甲醇转移缓冲液和飞克级化学发光剂信号均有助于小分子蛋白免疫印迹检测。 |
| 英文摘要: |
| Objective:Since the invention of the Western blotting method, it has been extensively applied as a protein qualitative
and semi-quantitative analysis method in modern biology research. In order to improve the efficiency of this method, adjustments of the
related experimental parameters were needed according to the characteristics of different proteins. Our purpose is to investigate the effects of different parameters of the Western blot in detecting small molecule proteins, thus optimize and get the ideal experimental conditions.Methods:Comparing the effects of different transfer voltages and times, different methanol contents of transfer buffer and different
chemiluminescence reagents of western blot technology.Results:The signal of small protein obtained at 20 V for 1 0 min was much higher than that transferred at 10 V for 25 min. The signal of the 20 % methanol in transfer buffer was significantly higher than that of the 0 %
methanol in transfer buffer. The effect of chemiluminescence reagent at femtogram level was superior to that of the nanogram level.Conclusion:Choosing high voltage and short time, choosing transfer buffer containing 20 % methanol and chemiluminescence reagent at
femtogram level could be helpful in the detection of small molecule proteins by using the western blot assay. |
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