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Tracing Pathway Activities with Kinase Inhibitors and Reverse Phase Protein Arrays by Jan van Oostrum; Claudio Calonder; Daniel Rechsteiner; Markus Ehrat; Jürgen Mestan; Doriano Fabbro; Hans Voshol is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Tracing Pathway Activities with Kinase Inhibitors and Reverse Phase Protein Arrays about?
## Abstract Controlling aberrant protein kinase activity is a promising strategy for a variety of diseases, particularly cancer. Hence, the development of kinase inhibitors is currently a focal point for pharmaceutical research. In this study we utilize a chip‐based reverse phase protein array (RPA) platform for profiling of kinase inhibitors in cell‐based assays. In combination with the planar wave‐guide technology the assay system has an absolute LOD down to the low zeptomole range. A431 cell lysates were analyzed for the activation state of key effectors in the epidermal growth factor (EGF) and insulin signaling pathways to validate this model for compound screening. A microtiter‐plate format for growing, treating, and lysing cells was shown to be suitable for this approach, establishing the value of the technology as a screening tool for characterization of large numbers of kinase inhibitors against a wide variety of cellular signaling pathways. Moreover, the reverse array format allows rapid development of site‐specific phosphorylation assays, since in contrast to ELISA type systems only a single antigen‐specific antibody is required.
Who reads Tracing Pathway Activities with Kinase Inhibitors and Reverse Phase Protein Arrays?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Jan van Oostrum; Claudio Calonder; Daniel Rechsteiner; Markus Ehrat; Jürgen Mestan; Doriano Fabbro; Hans Voshol
- Publisher
- John Wiley and Sons; Wiley (John Wiley & Sons); Wiley - VCH Verlag GmbH & CO. KGaA; Wiley (ISSN 1862-8346)
- Published
- 2009
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)