Solution Information | help | |
Enzyme: | Fibrinogen beta chain [164-491] | |
inhibitor: | BDBM25525 | |
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-TN7 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 (Tn7) to the fibrin proteolytic product D-Dimer and fragment E complex [DD(E )]" (AID 720628). 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 wells, followed | |
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