| Solution Information | help | |
| Enzyme: | Envelope glycoprotein gp160 | |
| inhibitor: | BDBM50618 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1) Env2.0 peptide Seq: Ac-MTWBEWDREIBNYTSLIC(FAM)-NH2 2) C18-Aib(FL) peptide Seq: bpy-GQAVEAQQHLLQLTVWGIKQLQARILAVEKK-NH2 3) Assay Buffer: 25 mM Tris-acetate pH 7.0, 0.01% Tween-20 4) The Fe(II) complex [Fe(II)(env2.0)3]2+ is prepared by addition of a one-third stoichiometry of freshly prepared ferrous ammonium sulfate to peptide in 25 mM Tris-acetate pH 7.0 Procedure Mix equal parts of the Env2.0/Fe complex and C18-Aib(FL) (60 uM) just prior to the assay 1) Using a Labcyte Echo, DMSO and test compounds are transferred to wells of a black, Corning 1536 well assay plate. DMSO only is transferred to columns 1-3 and 46-48(Control wells), while varying volumes of test compounds are transferred to columns 4-45 to achieve the desired test concentrations. Compounds are transferred from a 2 mM stock to give the stated final concentration. Test compound wells in the assay plate are back-filled with DMSO to equalize final assay concentrations. 2) Immediately following Echo T | |
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