Abstract
Protein microarrays are an ideal technology platform which allow for a robust and standardized profiling of the cellular proteome. Many cellular functions are not simply controlled by the presence of certain proteins, especially the propagation of external stimuli, which depend on transient post-translational modifications that determine whether a protein is in its active or inactive state. Thus, complex biological processes require the availability of a sound set of quantitative and time-resolved measurements to be understood. For this reason, new assay platforms which allow for the investigation of several proteins in parallel are necessary. The current best understood mode of cellular regulation occurs via phosphorylation and dephosphorylation processes, which are mediated via a large panel of kinases and phosphatases. The microspot immunoassay technique described here allows for an exact determination of several different phosphorylated proteins in parallel, as well as from small sample amounts, and is therefore an appropriate system to deepen the understanding of the complex regulatory networks implicated in health and disease.
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Acknowledgments
The authors would like to acknowledge S. Schumacher, M. Wosch, C. Becki, and C. Schmidt for their excellent technical support in the development of technically robust protocols. This work was supported by the German Federal Ministry for Education and Science in the framework of the Program for Medical Genome Research (grants 01GS0890 and 01GS0864), the Program for Medical Systems Biology (grant 0315396B), as well as the Helmholtz Systems Biology Initiative (SBCancer).
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Henjes, F., Götschel, F., Jöcker, A., Korf, U. (2011). Quantitative Analysis of Phosphoproteins Using Microspot Immunoassays. In: Korf, U. (eds) Protein Microarrays. Methods in Molecular Biology, vol 785. Humana Press. https://doi.org/10.1007/978-1-61779-286-1_13
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DOI: https://doi.org/10.1007/978-1-61779-286-1_13
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