We aim to quantitatively investigate and delineate the coordination of growth factor and hormone mediated phosphoregulation of intracellular signaling in breast cancer cell lines in a time-resolved fashion, and to subsequently validate newly identified key regulator proteins in tumor samples. To this end, protein microarray-based quantitative technologies have been established and are applied (Reverse Phase Protein Arrays - RPPA). We have set-up another experimental platform for the quantitative analysis of breast cancer patient plasma samples (Microspot Immuno Assays - MIA). Automated protocols have been developed to perform robust and artefact-free dynamic measurements in breast cancer cell lines relying on the use of a tissue-culture robot and allowing targeted perturbation experiments. The focus has been on analyzing intracellular signaling in response to receptor stimulation and to delineate the impact of targeted drugs in short-term (60 min) as well as long-term (30 h) measurements. To analyze data from large-scale protein microarray-experimentation, software tools tailored towards requirements for e.g., data normalization as well as data visualization have been developed.