| Solution Information | help | |
| Enzyme: | Neutrophil cytosol factor 1 [S99G] | |
| inhibitor: | BDBM38361 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Compounds marked as active in the primary screen were tested in this dose-response follow-up assay. Reagents: 1.Assay buffer: 20 mM Tris buffer, pH 7.5, 50 mM NaCl, and 0.01% NP40 2.p47phox protein 3.p22phox peptide with an N-terminal rhodamine label (p22-Rho) Procedure: 1. Make assay reaction buffer for uHTS that contains p47phox protein (100 nM, final concentration), p22-Rho peptide (15 nM, final concentration). 2. Dispense 8 uL of assay reaction buffer to 1536-well black assay plate. 3. Add library compound to each well and incubate plates at room temperature for 2 hr. 4. Record FP signals with an Envision Multilabel plate reader (Perkin Elmer Life Sciences. An excitation filter at 531 nm and emission filter at 595 nm are used with a 555 nm mirror. Data analysis: 1. FP signals are expressed as mP and recorded by Envision Multilabel plate reader. 2. Assay data are analyzed using CambridgeSoft and software from the R project for statistical computing. Percentage | |
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