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Journal of Biomolecular Screening
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Validation of a Cell-Based Screen for Insulin Receptor Modulators by Quantification of Insulin Receptor Phosphorylation

Lisa K. Minor

Johnson & Johnson Pharmaceutical Research and Development, L.L.C., Spring House, PA

Lori Westover

Johnson & Johnson Pharmaceutical Research and Development, L.L.C., Raritan, NJ

Yan Kong

Johnson & Johnson Pharmaceutical Research and Development, L.L.C., Raritan, NJ

Darpana Patel

Johnson & Johnson Pharmaceutical Research and Development, L.L.C., Raritan, NJ

Mary Jo Wildey

Johnson & Johnson Pharmaceutical Research and Development, L.L.C., Raritan, NJ

Bruce R. Conway

Johnson & Johnson Pharmaceutical Research and Development, L.L.C., Raritan, NJ

Keith T. Demarest

Johnson & Johnson Pharmaceutical Research and Development, L.L.C., Raritan, NJ

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.

Key Words: insulin • receptor phosphorylation • cell-based assays • fluorescence resonance energy transfer

Journal of Biomolecular Screening, Vol. 8, No. 4, 439-446 (2003)
DOI: 10.1177/1087057103255280
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