Open AccessAnalysis of Protein Expression in Cell Microarrays: A Tool for Antibody-based Proteomics
Author(s) -
AnnCatrin Andersson,
Sara Strömberg,
Helena Bäckvall,
Caroline Kampf,
Mathias Uhlén,
Kenneth Wester,
Fredrik Pontén
Publication year - 2006
Publication title -
journal of histochemistry and cytochemistry/the journal of histochemistry and cytochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.971
H-Index - 124
eISSN - 1551-5044
pISSN - 0022-1554
DOI - 10.1369/jhc.6a7001.2006
Subject(s) - tissue microarray , protein microarray , proteomics , microbiology and biotechnology , proteome , antibody , immunohistochemistry , agarose , cell , antibody microarray , protein array analysis , biology , cell culture , dna microarray , chemistry , computational biology , gene expression , bioinformatics , biochemistry , immunology , gene , genetics
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.