Figure 1.

High-throughput reverse phase protein array application to identify drug compound mechanism-of-action and compensatory response. (A) Schematic representation of RPPA workflow: Protein lysates are extracted from multiple samples and deposited across multiple sub-arrays on a microarray by high-precision printing. Validated antibodies are individually addressed to each array, followed by incubation with labeled secondary antibody detection reagent. Antibody binding to each sample is measured by an image-based microarray scanner followed by calculation of the relative abundance of each protein and post-translational modification across the sample set. (B) Quantification of the relative abundance of a set of 120 total protein and post-translational pathway markers following exposure to a small set of targeted therapies across dose-response. Un-annotated pathway data normalized to respective DMSO controls demonstrate significant enhancement of multiple pathway signaling events in addition to inhibition of targeted pathways.