| Solution Information | help | |
| Enzyme: | Nuclear hormone receptor family member daf-12 | |
| inhibitor: | BDBM114873 | |
| 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 dose response curves for compounds identified as activators of the nuclear receptor Daf-12 from S.stercoralis in a set of experiments entitled, "Luminescence-based cell-based primary high throughput screening assay to identify activators of the DAF-12 from the parasite S. stercoralis (ssDAF-12)" (AID 652126). Compounds are tested in triplicate using a 10-point, 1:3 dilution series starting at a nominal concentration of 68 uM. 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 8 ug of the ssDAF12-e | |
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