Receptor tyrosine kinase assays

Details Receptor tyrosine kinase assays Receptor tyrosine kinase assays

2009-08-06

2011-11-29

Woodward, Cherie M (Department: 1647)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

C435S007800, C435S007900

Reexamination Certificate

active

08067155

ABSTRACT:
Methods for detecting phosphorylation of receptor tyrosine kinases (“RTKs”) upon activation 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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