Validation of a cell-based screen for insulin receptor modulators by quantification of insulin receptor phosphorylation |
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Authors: | Minor Lisa K Westover Lori Kong Yan Patel Darpana Wildey Mary Jo Conway Bruce R Demarest Keith T |
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Affiliation: | Johnson & Johnson Pharmaceutical Research and Development, L L C, Spring House, PA 19477, USA. lminor@prdus.jnj.com |
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Abstract: | Stimulation of a cell with insulin initiates a signal transduction cascade that results in cellular activities that include phosphorylation of the receptor itself. Measurement of the degree of phosphorylation can serve as a marker for receptor activation. Receptor phosphorylation has been measured using Western blot analysis, which is very low throughput and not easily quantifiable. The goal of this project was to develop a cell-based assay to measure receptor phosphorylation in high throughput. This report describes a cell-based assay for insulin receptor phosphorylation that is robust and amenable to high-volume screening in a microwell format. |
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