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Originally published In Press as doi:10.1074/mcp.T600051-MCP200 on February 10, 2007.
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Molecular & Cellular Proteomics 6:939-947, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

Technology

Immunocell-array for Molecular Dissection of Multiple Signaling Pathways in Mammalian Cells*,S

Andrea Zanardi{ddagger}, Luca Giorgetti§,¶, Oronza A. Botrugno§,||, Saverio Minucci§, Paolo Milani, Pier Giuseppe Pelicci§,** and Roberta Carbone{ddagger},{ddagger}{ddagger}

From {ddagger} Tethis S.r.l., Via Russoli 3, 20143 Milan, Italy, § European Institute of Oncology (EIO), Via Ripamonti 435, 20141 Milan, Italy, CIMAINA, Centro Interdisciplinare Materiali e Interfacce Nanostrutturati, Via Celoria 16, 20133 Milan, Italy, and ** IFOM, Istituto Fondazione Italiana per la Ricerca sul Cancro (FIRC) di Oncologia Molecolare, Via Adamello 16, 20139 Milan, Italy

The knowledge of signaling pathways that are triggered by physiological and pathological conditions or drug treatment is essential for the comprehension of the biological events that regulate cellular responses. Recently novel platforms based on "reverse-phase protein arrays" have proven to be useful in the study of different pathways, but they still lack the possibility to detect events in the complexity of a cellular context. We developed an "immunocell-array" of cells on chip where, upon cell plating, growing, drug treatment, and fixation, by spotting specific antibodies we can detect the localization and state of hundreds of proteins involved in specific signaling pathways. By applying this technology to mammalian cells we analyzed signaling proteins involved in the response to DNA damage and identified a chromatin remodeling pathway following bleomycin treatment. We propose our technology as a new tool for the array-based multiplexed analysis of signaling pathways in drug response screening, for the proteomics of profiling patient cells, and ultimately for the high throughput screening of antibodies for immunofluorescence applications.

{ddagger}{ddagger} To whom correspondence should be addressed: Tethis S.r.l., Via Russoli 3, 20143 Milan, Italy. Tel.: 39-0236568349; Mobile: 39-392-9774561; Fax: 39-0236569183; E-mail: roberta.carbone{at}tethis-lab.com


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