| Solution Information | help | |
| Enzyme: | Tegument protein VP16 | |
| inhibitor: | BDBM76151 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine whether the hits identified as active in a set of experiments entitled, Luminescence-based cell-based primary high throughput screening assay to identify antagonists of COUP-TFII (NR2F2) (AID 686940), are assay artifacts affecting luciferase activity, cell viability or the transcription/translation machinery in a non-selective manner. Protocol Summary: HEK293 cells were routinely cultured in T-175 flasks containing 25 mL of DMEM media supplemented with 10% v/v fetal bovine serum and 1% v/v antibiotic-antimycotic mix at 37 C, 5% CO2 and 95% relative humidity (RH). The day prior to run the assay, the HEK293 cells were harvested using 5 mL of TrypLE reagents and seeded in fresh media at a density of 10 million cells per T175 flask. The following day, cells were transfected with 1 mL of serum-free OptiMEM containing 9.9 ug of the plasmid encoding VP16 (pAB-Gal4-VP16), 9.9 ug of the p17 mer x4-TK-Luc luciferase reporter plasmid, | |
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