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Analysis of protein expression in cell microarrays: A tool for antibody-based proteomics
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2006 (English)In: Journal of Histochemistry and Cytochemistry, ISSN 0022-1554, E-ISSN 1551-5044, Vol. 54, no 12, 1413-1423 p.Article in journal (Refereed) Published
Abstract [en]

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 TIVIA 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.

Place, publisher, year, edition, pages
2006. Vol. 54, no 12, 1413-1423 p.
Keyword [en]
immunohistochemistry, cell line, tissue microarray, affinity proteomics, antibody-based proteomics, anticancer drug screen, high-throughput, tissue microarrays, cultured-cells, lines, cancer, immunohistochemistry, quantification, establishment, technology
Identifiers
URN: urn:nbn:se:kth:diva-16131DOI: 10.1369/jhc.6A7001.2006ISI: 000242056300010Scopus ID: 2-s2.0-33751203277OAI: oai:DiVA.org:kth-16131DiVA: diva2:334173
Note
QC 20100525Available from: 2010-08-05 Created: 2010-08-05 Last updated: 2017-12-12Bibliographically approved

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Uhlén, Mathias

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