| Solution Information | help | |
| Enzyme: | Glycoprotein 42 | |
| inhibitor: | BDBM31734 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Positives from the primary screen, AID 1085, were selected for testing in this confirmatory assay. Reagents: 1.Assay buffer: 20 mM Tris buffer, pH 7.5, 50 mM NaCl, and 0.01% NP40 2.gH/gL protein 3.gp42-36-81 peptide with an N-terminal fluorescein label (gp42-Flu) Procedure: 1. Make assay reaction buffer for uHTS that contains gH/gL protein (2 nM, final concentration), gp42-Flu peptide (2 nM, final concentration). 2. Dispense 4.5 uL of assay reaction buffer to 1536-well black assay plate. 3. Add appropriate amount of compound to each well to reach assay concentration and incubate plates at room temperature for 2 hr. 4. Record FP signals with an Envision Multilabel plate reader (Perkin Elmer Life Sciences. Data analysis: 1.FP signals are expressed as mP and recorded by Envision Multilabel plate reader. 2.Assay data are analyzed using CambridgeSoft. Percentage of inhibition is calculated with the following equation based on data from each plate. % of Inhibition = 100 - ((FPco | |
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