Abstract
In most hospitals around the world FFPE (formalin fixed, paraffin embedded) tissues have been used for diagnosis and have subsequently been archived since decades. This has lead to a sizeable pool of this kind of tissues. Till quite recently it was not possible to use this congeries of samples for protein analysis, but now several groups described successful protein extraction from FFPE tissues. In this chapter, we describe a protein extraction protocol established in our laboratory combined with the use of reverse phase protein microarray.
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Becker, K. F., Schott, C., Hipp, S., Metzger, V., Porschewski, P., Beck, R., Nahrig, J., Becker, I. and Hofler, H. (2007) Quantitative protein analysis from formalin-fixed tissues: implications for translational clinical research and nanoscale molecular diagnosis. J Pathol 211, 370–8.
Addis, M. F., Tanca, A., Pagnozzi, D., Crobu, S., Fanciulli, G., Cossu-Rocca, P., and Uzzau, S. (2009) Generation of high-quality protein extracts from formalin-fixed, paraffin-embedded tissues. Proteomics 9, 3815–23.
Chu, W.S., Liang, Q., Liu, J., Wei, MQ., Winters, M., Liotta, L., Sandberg, G., and Gong, M. (2005) A nondestructive molecule extraction method allowing morphological and molecular analyses using a single tissue section. Lab Invest 85, 1416–28.
Chung, J., Lee, S. J., Kris, Y., Braunschweig, T., Traicoff, J. L., and Hewitt, S. M. (2008) A well-based reverse-phase protein array applicable to extracts from formalin-fixed paraffin-embedded tissue. Proteomics Clin App 2, 1539–47.
Ikeda, K., Monden, T., Kanoh, T., Tsujie, M., Izawa, H., Haba, A., Ohnishi, T., Sekimoto, M., Tomita, N., Shiozaki, H., and Monden, M. (1998) Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tissue sections. J Histochem Cytochem 46, 397–403.
Nirmalan, N. J., Harnden, P., Selby, P. J., and Banks, R. E. (2009) Development and validation of a novel protein extraction methodology for quantitation of protein expression in formalin-fixed paraffin-embedded tissues using western blotting. J Pathol 217, 497–506.
Shi, S. R., Liu, C., Balgley, B. M., Lee, C., and Taylor, C. R. (2006) Protein extraction from formalin-fixed, paraffin-embedded tissue sections: quality evaluation by mass spectrometry. J Histochem Cytochem 54, 739–43.
Becker, K. F., Mack, H., Schott, C., Hipp, S., Rappl, A., Piontek, G., and Höfler, H. (2008) Extraction of phosphorylated proteins from formalin-fixed cancer cells and tissues. TOPATJ 2, 44–52.
Becker, K. F., Schott, C., Becker, I., and Höfler, H. (2008) Guided protein extraction from formalin-fixed tissues for quantitative multiplex analysis avoids detrimental effects of histological stains. Proteomics Clin Appl. 2, 737–43.
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Wolff, C., Schott, C., Malinowsky, K., Berg, D., Becker, KF. (2011). Producing Reverse Phase Protein Microarrays from Formalin-Fixed Tissues. In: Korf, U. (eds) Protein Microarrays. Methods in Molecular Biology, vol 785. Humana Press. https://doi.org/10.1007/978-1-61779-286-1_9
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DOI: https://doi.org/10.1007/978-1-61779-286-1_9
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Online ISBN: 978-1-61779-286-1
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