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采用多条带蛋白质免疫印迹法以增加定量数据输出。

Multistrip western blotting to increase quantitative data output.

作者信息

Kiyatkin Anatoly, Aksamitiene Edita

机构信息

Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, JAH 233, 1020 Locust Street, Philadelphia, PA, 19107, USA.

出版信息

Methods Mol Biol. 2009;536:149-61. doi: 10.1007/978-1-59745-542-8_17.

Abstract

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.

摘要

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。

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本文引用的文献

1
Multistrip Western blotting to increase quantitative data output.
Electrophoresis. 2007 Sep;28(18):3163-73. doi: 10.1002/elps.200700002.
2
Strategies for plasma proteomic profiling of cancers.
Proteomics. 2006 Oct;6(20):5662-73. doi: 10.1002/pmic.200600331.
3
Quantitative data generation for systems biology: the impact of randomisation, calibrators and normalisers.
Syst Biol (Stevenage). 2005 Dec;152(4):193-200. doi: 10.1049/ip-syb:20050044.
4
A novel method of loading samples onto mini-gels for SDS-PAGE: increased sensitivity and Western blots using sub-microgram quantities of protein.
J Neurosci Methods. 2006 Dec 15;158(2):224-33. doi: 10.1016/j.jneumeth.2006.06.002. Epub 2006 Jul 10.
5
Proteomic biomarkers for autoimmune disease.
Proteomics. 2006 Jul;6(14):4100-5. doi: 10.1002/pmic.200600017.
6
Western blotting.
Methods. 2006 Apr;38(4):283-93. doi: 10.1016/j.ymeth.2005.11.007.
7
Cell-signalling dynamics in time and space.
Nat Rev Mol Cell Biol. 2006 Mar;7(3):165-76. doi: 10.1038/nrm1838.
9
Tyrosine kinases as targets for cancer therapy.
N Engl J Med. 2005 Jul 14;353(2):172-87. doi: 10.1056/NEJMra044389.
10
Dual specificity protein phosphatases: therapeutic targets for cancer and Alzheimer's disease.
Annu Rev Pharmacol Toxicol. 2005;45:725-50. doi: 10.1146/annurev.pharmtox.45.120403.100040.

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