Solution Information | help | |
Enzyme: | Focal adhesion kinase 1 | |
inhibitor: | BDBM31698 | |
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 dose response curves for 210 available out of 212 compounds identified from a previously described set of experiments entitled "Confirmation biochemical assay for inhibitors of Focal Adhesion Kinase (FAK)" (PubChem AID = 794). Each compound was assayed in triplicate using ten 1:3 serial dilutions, starting at a nominal test concentration of 70 micromolar. As with the primary and confirmation screens, a homogeneous time-resolved fluorescence resonance energy transfer (TR-FRET) method to monitor the phosphorylation of a poly Glu-Tyr (poly-GT) substrate by FAK was used to screen for inhibitors. The principle of the assay is similar to other kinase detection systems utilizing TR-FRET, except this method utilizes a FRET acceptor dye which is coupled directly to the substrate. The level of substrate phosphorylation by FAK activity is detected with a specific europium-labeled anti-phospho-peptide antibody (Eu-Ab), which serves as t | |
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