| Autor: |
Robert A. Davey, Lisa A. Elferink, Ivana Gaziova |
| Rok vydání: |
2014 |
| Předmět: |
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| Zdroj: |
Membrane Trafficking ISBN: 9781493923083 |
| DOI: |
10.1007/978-1-4939-2309-0_26 |
| Popis: |
The tyrosine kinase MET, a receptor for hepatocyte growth factor, is a key regulator for normal development and organ renewal via stem cell maintenance. Dysregulated MET signaling contributes to tumor progression and metastasis and is considered a potent therapeutic target for a growing number of malignancies. Toward that goal it is critical to develop high-throughput assays to identify candidate regulators for the termination of MET signaling. We describe here a rapid and efficient method for identifying cellular factors required for MET ubiquitination, which utilizes high-throughput RNA interference screening (HT-siRNA) with a receptor internalization assay and an In-Cell ELISA in a 96-well format. The assay is amenable to a large array of cell surface proteins as well as genome-wide siRNA libraries, with high signal-to-background ratio and low well-to-well variability. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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