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Methods Mol Biol. 2004;264:1-13.

Protein arrays from cDNA expression libraries.

Author information

1
Department of Vertebrate Genomics, Max Planck Institute of Molecular Genetics, Berlin, Germany.

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.

PMID:
15020775
DOI:
10.1385/1-59259-759-9:001
[Indexed for MEDLINE]

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