Unique, whole cell Assay platform for Screening of Receptor Tyrosine Kinase Activity
The PathHunter™ technology is an adaptation of Enzyme Fragment Complementation (EFC) that provides a novel, generic cell-based assay format for detecting protein-protein interactions. Receptor Tyrosine Kinases are implicated in disease states such as cancer, metabolic disease, inflammatory disease, cardiovascular and many other different disease areas. In this PathHunter assay approach, a small peptide epitope (ProLink) is expressed recombinantly on the intracellular C-terminus of the Receptor Tyrosine Kinase. One of the many different partner proteins containing SH2 domains (such as SHC1, SHC2, Grb2, PTPN6, PLCG1 or PLCG2) is co-expressed with a larger sequence, termed enzyme acceptor (EA). Ligand induced activation of the receptor causes either homo- or hetero-dimerization of the receptor resulting in cross-phosphorylation. The SH2-EA fusion protein then binds the phosphorylated receptor forcing complementation of the PK and EA fragment. This interaction generates an active beta-galactosidase enzyme, which is detected using a chemiluminescent substrate.
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