RECEPTOR TYROSINE KINASE ASSAYS
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United States of America Patent
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Issued Date -
Feb 16, 2012
app pub date -
Oct 25, 2011
filing date -
Aug 18, 2008
priority date (Note) -
Abandoned
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Abstract
Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation and the modulation of activation by a candidate compound are provided. The method employs cells comprising two fusion products: (1) an RTK fused to a small fragment of β-galactosidase and (2) a phosphotyrosine binding peptide fused to the large fragment of β-galactosidase, where the 2 fragments weakly complex to form an active enzyme, and optionally a construct for a cytosolic RTK phosphorylating kinase, when the RTK does not autophosphoryate. To detect phosphorylation a β-galactosidase substrate is added to the cells, whereby product formation indicates the occurrence of phosphorylation.
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