Abstract
Tissue interstitial fluid (TIF) forms the interface between circulating body fluids and intracellular fluid. Pathological alterations of liver cells could be reflected in TIF, making it a promising source of liver disease biomarkers. Here, we introduce the method of extracting TIF from mouse liver. We confirm the absence of cellular protein contamination by western blot using organelle specific protein markers including lamin B, cytochrome C, and flotillin-1. Using two-dimensional differential in-gel electrophoresis (2D-DIGE), we demonstrate different profiling patterns among TIFs of liver tissue and hepatocytes. The high solubility and even distribution of liver TIF supports its suitability for proteome analysis. Based on the method introduced here, liver TIF in animal models and clinical samples could be extracted and analyzed for further application in liver disease biomarker discovery.
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Acknowledgments
We thank Dr. Celis Julio E. for the constructive discussion with him on the extraction protocol. Also, we thank Drs. Yunwei Hao, Lihai Guo, Ping Wan, Yuanbiao Guo, and Xinyu Deng for their help and advices. This work was partially supported by Chinese State High-tech Program (2006AA02A308), Chinese State Key Projects for Basic Research (Nos. 2006CB910801, 2006CB910401, 2006CB910602, 2010CB912700 and 2006CB503910), National Natural Science Foundation of China (30700988, 30700356, 30972909), Chinese State Key Project Specialized for Infectious Diseases (2008ZX10002-016, 2009ZX10004-103, 2009ZX09301-002), International Scientific Collaboration Program (2009DFB33070, 2008GR0763) and supported by the State Key Laboratory of Proteomics (SKLP-K200801, SKLP-Y200801, SKLP-O200801) in part with permission from Sun et al. (2010) (4). Copyright 2010 American Chemical Society.
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Sun, W., Jiang, Y., He, F. (2011). Extraction and Proteome Analysis of Liver Tissue Interstitial Fluid. In: Simpson, R., Greening, D. (eds) Serum/Plasma Proteomics. Methods in Molecular Biology, vol 728. Humana Press. https://doi.org/10.1007/978-1-61779-068-3_16
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DOI: https://doi.org/10.1007/978-1-61779-068-3_16
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Online ISBN: 978-1-61779-068-3
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