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Microwestern Assay

Microwestern arrays are a systems biology approach for quantitative multiplex analysis of cell signaling pathways and can be performed on the Odyssey® CLx or Odyssey Classic Imaging System.

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

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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 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® Classic 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 approppriate computational modeling methods.

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Effect of complementary pathway blockade on efficacy of combination enzastaurin and rapamycin.
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Systematic Discovery of TLR Signaling Components Delineates Viral-Sensing Circuits
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Ciaccio, M.F., et al.
Systems Analysis of EGF Receptor Signaling Dynamics with Micro-Western Arrays
Nat. Methods, Feb 2010; 7: 148–155.

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