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Assay Method Information

Assay Name:  Biological Assay
Description:  TrkA functional activity was measured using a DiscoverX PathHunter assay. In this assay, U2OS cells express the human TrkA receptor as a fusion with the weakly complementing fragment of B-galactosidase, which DiscoverX calls Prolink (PK) ; additionally, Shc1 is fused with a larger fragment, which is called Enzyme Acceptor (EA) . Activation of the TrkA receptor, upon NGF addition, results in the kinase domain being phosphorylated, resulting in subsequent recruitment of Shc1-EA protein. That recruitment results in an active B-galactosidase enzyme that is detected by addition of a chemiluminescent substrate. The human p75NTR protein was also expressed as a co-receptor for NGF.
Affinity data for this assay
 

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Last update November 1, 2007
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