| Solution Information | help | |
| Enzyme: | fMet-Leu-Phe receptor | |
| inhibitor: | BDBM37628 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | For assay performance, additions to wells were in sequence as follows: 1) test compounds and control reagents (5 microL/well); 2) a combination of FPR- and FPRL1-expressing cell lines (10^7/mL each, 5 microL/well); 3) (after 30 min, 4 degrees C incubation) fluorescent peptide (5 microL/well). After an additional 45 min, 4 degrees C incubation, plates were immediately analyzed by flow cytometry. The assay response range was defined by replicate control wells containing unlabeled receptor-blocking peptide (positive control) or buffer (negative control). fMLFF (4Pep) was used as the FPR-blocking peptide, unlabeled WPep as the FPRL1-blocking peptide. The assay was homogeneous in that cells, compounds and fluorescent peptide were added in sequence and the wells subsequently analyzed without intervening wash steps. The HyperCyt high throughput flow cytometry platform (Kuckuck et al., 2001; Ramirez et al., 2003) was used to sequentially sample cells from wells of 384-well microplates (2 mi | |
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