| Assay Method Information | |
| | Inhibition of C5a-C5aR Binding Determined by Cell Migration Assays |
| Description: | Migration assay was performed by using polycarbonate membrane with 3.0 m pore (Corning). U937/C5aR cells were collected and washed twice by PBS; cells were suspended in Hank's balanced salt solution (HBSS)+1% FBS buffer at the density of 6×106 cells/ml. Cells were premixed with compound and added to insert well, ligand human C5a and compound were added to bottom well in order, gently mix, incubate for 30 min at 37° C., 500 CO2. Put insert plate into bottom well, migrate for 180 min at 37° C., 500 CO2. Gently remove the insert well, add 50 μl CellTiter-Glo (Promega), gently shaking for 5 min at room temperature, transfer 150 μl mixture to black plate and read luminescence intensity byConfidential Draft microplate reader (BioTek). |
| Affinity data for this assay | |
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