Table 2.
Protocol of the Multiplexed Calcium Assay in 1,536-well Format
| Step | Parameter | Value | Description |
|---|---|---|---|
| 1 | Cells | 3 μL | 1,500 cells/well |
| 2 | Incubation | 16–20 h | 37°C, 5% CO2 |
| 3 | Reagent | 3 μL | No-wash calcium dye |
| 4 | Incubation | 60 min | 37°C, 5% CO2 |
| 5 | Incubation | 10 min | Room temperature |
| 6 | Detection | Basal signal | Read for 10 s in the FDSS7000 |
| 7 | Compounds | 20 nL | Added by pin tool in the instrument |
| 8 | Detection | Agonist response | Read for 3.5 min |
| 9 | Reagent | 2 μL | Add EC90 of agonist |
| 10 | Detection | Antagonist response | Read for 140 s |