| Solution Information | help | |
| Enzyme: | Nuclear hormone receptor family member daf-12 | |
| inhibitor: | BDBM1735 | |
| 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 the nuclear receptor Daf-12 from H.contortus agonists dose response curves for compounds that confirmed activity in a set of previous experiments entitled: "Luminescence-based cell-based primary high throughput screening assay to identify activators of the DAF-12 from the parasite H. contortus (hcDAF-12)" (AID 652067). In this assay, HEK293 cells are co-transfected with a DAF12-responsive reporter plasmid (lit1-tk-luc) and expression vector encoding hcDAF12. The ability of compounds to increase transcriptional activity is assessed by measuring luciferase expression from the reporter gene plasmid. 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% relati | |
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