| Assay Method Information | |
| | Calcium Mobilization |
| Description: | U937/C5aR cells or HEK293/C5aR cells were washed by PBS and suspended in growth media at the density of 1 106 cells/ml. Seed 2 cell suspension to the 384-well plate and culture for overnight. Transfer 250 nl compound solution to the cell plate using Echo, incubate for 60 min. Cells were added with Fluo-4 Direct dye and incubate for 50 min at 37 C. 5% CO2 and 10 m at room temperature. Place the cell plate into FLIPRTETRA (Molecular Devices). Transfer 10 μl of 5-fold EC80 concentrations of agonist human C5a to the cell plates. Read fluorescence signal, data was calculated by GraphPad Prism. |
| Affinity data for this assay | |
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