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1: J Histochem Cytochem. 2006 Dec;54(12):1413-23. Epub 2006 Sep 6.
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Analysis of protein expression in cell microarrays: a tool for antibody-based proteomics.

Andersson AC, Strömberg S, Bäckvall H, Kampf C, Uhlen M, Wester K, Pontén F.

Department of Genetics and Pathology, Rudbeck Laboratory, University Hospital, Uppsala, Sweden.

Tissue microarray (TMA) technology provides a possibility to explore protein expression patterns in a multitude of normal and disease tissues in a high-throughput setting. Although TMAs have been used for analysis of tissue samples, robust methods for studying in vitro cultured cell lines and cell aspirates in a TMA format have been lacking. We have adopted a technique to homogeneously distribute cells in an agarose gel matrix, creating an artificial tissue. This enables simultaneous profiling of protein expression in suspension- and adherent-grown cell samples assembled in a microarray. In addition, the present study provides an optimized strategy for the basic laboratory steps to efficiently produce TMAs. Presented modifications resulted in an improved quality of specimens and a higher section yield compared with standard TMA production protocols. Sections from the generated cell TMAs were tested for immunohistochemical staining properties using 20 well-characterized antibodies. Comparison of immunoreactivity in cultured dispersed cells and corresponding cells in tissue samples showed congruent results for all tested antibodies. We conclude that a modified TMA technique, including cell samples, provides a valuable tool for high-throughput analysis of protein expression, and that this technique can be used for global approaches to explore the human proteome.

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PMID: 16957166 [PubMed - indexed for MEDLINE]