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Systems analysis of EGF receptor signaling
dynamics with microwestern arrays

Mark F Ciaccio, Joel P Wagner, Chih-Pin Chuu, Douglas A Lauffenburger & Richard B Jones

The Ben May Department for Cancer Research and the Institute for Genomics and Systems Biology, The University of Chicago, Chicago, Illinois. Center for Cell Decision Processes and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, Present address: Institute of Cellular and Systems Medicine, National Health Research Institutes, Miaoli County, Taiwan.

Nature Methods 7: 148 (2010)

Ciaccio et al. developed a method for doing higher throughput Western blotting using using 200-fold less protein sample and antibody called MicroWestern Arrays (MWAs). MWAs have the scalability of reverse phase arrays yet they retain the vital attributes of western blots where signals that can be related to protein size. The method should be use­ful for analysis of proteins from cell lines and tissues in an analogous manner to spotted DNA microarrays for interrogation but with the user's choice of antibodies. Protein samples are spotted on acrylamide slabs in a 96 well array, electophoresed, transferred to a membrane, placed in 96-well gasket device and imaged on the Odyssey Infrared Imaging System. The ability to obtain information regarding hundreds of proteins with the MWAs should allow advances in our understanding of cell context–specific networks underlying human disease when combined with appro­priate computational modeling methods.

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