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| Solution Information | help | |
| Enzyme: | Protein-tyrosine kinase 2-beta | |
| inhibitor: | BDBM41000 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine whether compounds identified as active in a previous set of experiments entitled, "Primary biochemical high-throughput screening assay for inhibitors of Focal Adhesion Kinase (FAK)" (PubChem AID 727), that confirmed activity in a set of experiments entitled, "Confirmation biochemical assay for inhibitors of Focal Adhesion Kinase (FAK)" (PubChem AID 794), and that were active in a set of experiments entitled, "Dose-response biochemical assay for inhibitors of Focal Adhesion Kinase (FAK)" (PubChem AID 810), were nonselective inhibitors due to inhibition of PYK2. In this time-resolved fluorescence resonance energy transfer (TR-FRET)-based assay, PYK2-mediated phosphorylation of a poly Glu-Tyr (poly-GT) substrate is monitored using a FRET acceptor dye which is coupled directly to the substrate. Substrate phosphorylation by PYK2 is detected with a specific europium-labeled anti-phospho-tyrosine PY20 antibody (Eu-Ab), which serves | |
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