Kiyatkin Anatoly, Aksamitiene Edita
Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, JAH 233, 1020 Locust Street, Philadelphia, PA, 19107, USA.
Methods Mol Biol. 2009;536:149-61. doi: 10.1007/978-1-59745-542-8_17.
The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single membrane sheet. In comparison with the conventional technique, Multistrip western blotting increases the data output per single blotting cycle up to tenfold, allows concurrent monitoring of up to nine different proteins from the same loading of the sample, and substantially improves the data accuracy by reducing immunoblotting-derived signal errors. This approach enables statistically reliable comparison of different or repeated sets of data, and therefore is beneficial to apply in biomedical diagnostics, systems biology, and cell signaling research.
蛋白质丰度和修饰状态的定性和定量测量对于理解其在各种细胞过程中的功能至关重要。典型的蛋白质印迹法虽然灵敏,但容易产生大量误差,且不易适应高通量技术。多通道蛋白质印迹法是一种改良的免疫印迹程序,基于将蛋白质从多条聚丙烯酰胺凝胶同时电泳转移到单个膜片上。与传统技术相比,多通道蛋白质印迹法将每个单次印迹循环的数据输出提高了十倍,能够同时监测来自相同样品上样量的多达九种不同蛋白质,并通过减少免疫印迹产生的信号误差大幅提高数据准确性。这种方法能够对不同或重复的数据集进行统计学上可靠的比较,因此有利于应用于生物医学诊断、系统生物学和细胞信号研究。