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October 30, 2012
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A high-content screening method for the identification of novel signaling competent transmembrane receptors is described. This method is amenable to large-scale automation and allows predictions about in vivo protein binding and the sub-cellular localization of protein complexes in mammalian cells.
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High-content ImagingGrb2 Signaling ComplexesExpression CloningSignal TransductionGrowth Factor ReceptorsDownstream Signaling CascadesMitogenic SignalingSH2 DomainSH3 DomainGTPase Son-Of-Sevenless (SOS)Phosphorylated Tyrosine ResiduesSOS-Ras-MAP Kinase Signaling CascadePositive And Negative Regulator Of SignalingReceptor EndocytosisMicroscopy AssayPlasma Membrane ReceptorsEpidermal Growth Factor Receptor (EGFR)GFP-Grb2
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Freeman, J., Kriston-Vizi, J., Seed, B., Ketteler, R. A High-content Imaging Workflow to Study Grb2 Signaling Complexes by Expression Cloning. J. Vis. Exp. (68), e4382, doi:10.3791/4382 (2012).
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