Abstract
This chapter describes the production of a cDNA expression library from human fetal brain, the construction of a high-density protein array from such a library, and two applications to screen the array for binding proteins. After producing the library and decollating the expression clones, one can pick thousands of expression clones with a laboratory robot and can deposit them into microtiter plates in an ordered manner. Such ordered clone libraries are the starting material for the construction of a high-density protein array. This array is constructed by spotting the expression clones onto a protein-binding membrane. Following cell growth and induction of protein expression on the membrane, the cell spots are lysed and their recombinant protein immobilized on the membrane. The so-constructed array carries thousands of proteins without the need to clone, express, and spot individual proteins. Such arrays allow one to screen for numerous protein functions in a high-throughput manner.
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© 2004 Humana Press Inc., Totowa, NJ
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Weiner, H., Faupel, T., Büssow, K. (2004). Protein Arrays From cDNA Expression Libraries. In: Fung, E.T. (eds) Protein Arrays. Methods in Molecular Biology, vol 264. Humana Press. https://doi.org/10.1385/1-59259-759-9:001
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DOI: https://doi.org/10.1385/1-59259-759-9:001
Publisher Name: Humana Press
Print ISBN: 978-1-58829-255-1
Online ISBN: 978-1-59259-759-8
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