Probe Reports from the NIH Molecular Libraries Program [Internet].
Bethesda (MD): National Center for Biotechnology Information (US); 2010-.
NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.
Table 3Stepwise protocol of the PGE2 HTRF assay using A549 cells in 1536-well format
| Sequence | Parameter | Value | Description |
|---|---|---|---|
| 1 | Reagent | 5 μL | 2,500 cells/well (A549 or media) |
| 2 | Compound | 23 nL | Compounds and controls |
| 3 | Time | Varied | Incubation at 37 ºC, 5% CO2, 4, 8, 12, and 16 hours |
| 4 | Reagent | 2.5 μL | PGE2-d2 |
| 5 | Reagent | 2.5 μL | anti-PGE2-cryptate |
| 6 | Time | 15 sec | Centrifuged at 1,500 RPM |
| 7 | Time | 5 hr | Incubation at room temperature, protected from light |
| 8 | Detector | HTRF | Envision (320 nm, 615 and 665 nm emission) |