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| Solution Information | help | |
| Enzyme: | Fibrinogen beta chain [164-491] | |
| inhibitor: | BDBM114655 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this biochemical assay is to determine Fibrin-TN6 inhibitory dose response curves for compounds that confirmed activity in a set of previous experiments entitled, "Fluorescence polarization-based biochemical high throughput confirmation assay to identify inhibitors that disrupt the binding of a cyclic peptide (Tn6) to the fibrin proteolytic product D-Dimer and fragment E complex [DD(E )]" (AID 720631). As designed, test compounds that bind to Fibrin-DD(E) will prevent Fibrin-DD(E)-probe interactions, thereby increasing the proportion of free (unbound) fluorescent probe in the well, leading to low fluorescence polarization. Compounds are tested in triplicate using a 10-point 1:3 dilution series starting at a maximum nomimal test concentration of 83.3 uM. Protocol Summary: Displacement assay starts by the addition of Fibrin - DD(E) (final concentration 2uM) in assay buffer (50mM Tris base, 100 mM NaCl, 2 mM CaCl2, and 0.01% Triton X-100, pH 7.8) to all | |
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