**10. References**


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TLCA: Taurolithocholic acid UDCA: Ursodeoxycholicacid

**Acknowledgement** 

Nacional del Sur, PGI 24/B152.

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UV: Ultraviolet

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**Chapter 3** 

© 2012 Van Chi and Dung, licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2012 Van Chi and Dung, licensee InTech. This is a paper distributed under the terms of the Creative Commons

**2D-NanoLC-ESI-MS/MS for** 

Phan Van Chi and Nguyen Tien Dung

http://dx.doi.org/10.5772/47479

**1. Introduction** 

demonstrated [16, 30].

Additional information is available at the end of the chapter

**Separation and Identification of** 

**Mouse Brain Membrane Proteins** 

Comprehensive proteomics analysis has the potential to provide new knowledge on cellular responses in development, aging, drug action, environmental stress, and disease pathogenesis (carcinogenesis, cardiovascular disease, etc). However, the separation and identification of proteomes/proteins is a challenging task due to their heterogeneous constituents or complex structures and closely related physico-chemical behaviors. It is clear that the combination of many analytical techniques is necessary to fulfill this complex task. At the start of proteomics research, two-dimensional electrophoresis (2DE) was routinely used to separate complex proteomic sample because of its high resolving power. In this technique, proteins are separated in a two-step process (two dimensions) based on their different physical properties. The first dimension is isoelectrofocusing in which proteins are separated based on their isoelectric points (pI, the pH where a *protein's net charge* is zero) using immobilized pH-gradient strips. Proteins then are separated according to their mass using sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension. With 2DE, thousands of proteins can be detected in a single experiment depending on the used staining techniques (Coomassie blue, silver, fluorescent dyes staining) [11]. Mass spectrometry (MS), using either electrospray ionization (ESI) or matrixassisted laser desorption/ionization (MALDI), is the key technology for the identification of protein spots including membrane proteins, for which differential expression has been

2DE, however, has some major drawbacks/disadvantages. It is time-consuming, difficult to reproduce and automation is hard to achieve. Furthermore, 2DE faces with many difficulties in analyzing several groups of proteins, such as low-abundance proteins, hydrophobic


**Chapter 3** 
